RESUMO
Understanding how nutrient stress impacts plant growth is fundamentally important to the development of approaches to improve crop production under nutrient limitation. Here we applied single-cell RNA sequencing to shoot apices of Pisum sativum grown under boron (B) deficiency. We identified up to 15 cell clusters based on the clustering of gene expression profiles and verified cell identity with cell-type-specific marker gene expression. Different cell types responded differently to B deficiency. Specifically, the expression of photosynthetic genes in mesophyll cells (MCs) was down-regulated by B deficiency, consistent with impaired photosynthetic rate. Furthermore, the down-regulation of stomatal development genes in guard cells, including homologs of MUTE and TOO MANY MOUTHS, correlated with a decrease in stomatal density under B deficiency. We also constructed the developmental trajectory of the shoot apical meristem (SAM) cells and a transcription factor interaction network. The developmental progression of SAM to MC was characterized by up-regulation of genes encoding histones and chromatin assembly and remodeling proteins including homologs of FASCIATA1 (FAS1) and SWITCH DEFECTIVE/SUCROSE NON-FERMENTABLE (SWI/SNF) complex. However, B deficiency suppressed their expression, which helps to explain impaired SAM development under B deficiency. These results represent a major advance over bulk-tissue RNA-seq analysis in which cell-type-specific responses are lost and hence important physiological responses to B deficiency are missed. The reported findings reveal strategies by which plants adapt to B deficiency thus offering breeders a set of specific targets for genetic improvement. The reported approach and resources have potential applications well beyond P. sativum species and could be applied to various legumes to improve their adaptability to multiple nutrient or abiotic stresses.
Assuntos
Boro , Pisum sativum , Pisum sativum/genética , Boro/metabolismo , Meristema/genética , Fatores de Transcrição/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas/genéticaRESUMO
Boric acid is a vital micronutrient in animals; however, excess amounts are toxic to them. Little is known about whole-body boric acid homeostasis in animals. Seawater (SW) contains 0.4 mM boric acid, and since marine fish drink SW, their urinary system was used here as a model of the boric acid excretion system. We determined that the bladder urine of a euryhaline pufferfish (river pufferfish, Takifugu obscurus) acclimated to fresh water and SW contained 0.020 and 19 mM of boric acid, respectively (a 950-fold difference), indicating the presence of a powerful excretory renal system for boric acid. Slc4a11 is a potential animal homolog of the plant boron transporter BOR1; however, mammalian Slc4a11 mediates H+ (OH-) conductance but does not transport boric acid. We found that renal expression of the pufferfish paralog of Slc4a11, Slc4a11A, was markedly induced after transfer from fresh water to SW, and Slc4a11A was localized to the apical membrane of kidney tubules. When pufferfish Slc4a11A was expressed in Xenopus oocytes, exposure to media containing boric acid and a voltage clamp elicited whole-cell outward currents, a marked increase in pHi, and increased boron content. In addition, the activity of Slc4a11A was independent of extracellular Na+. These results indicate that pufferfish Slc4a11A is an electrogenic boric acid transporter that functions as a B(OH)4- uniporter, B(OH)3-OH- cotransporter, or B(OH)3/H+ exchanger. These observations suggest that Slc4a11A is involved in the kidney tubular secretion of boric acid in SW fish, probably induced by the negative membrane potential and low pH of urine.
Assuntos
Boro , Rim , Proteínas de Membrana Transportadoras , Animais , Boro/metabolismo , Rim/metabolismo , Proteínas de Membrana Transportadoras/metabolismo , Água do Mar , Peixes , TakifuguRESUMO
The mechanistic basis by which boron (B) deprivation inhibits root growth via the mediation of root apical auxin transport and distribution remains elusive. This study showed that B deprivation repressed root growth of wild-type Arabidopsis seedlings, which was related to higher auxin accumulation (observed with DII-VENUS and DR5-GFP lines) in B-deprived roots. Boron deprivation elevated the auxin content in the root apex, coinciding with upregulation of the expression levels of auxin biosynthesis-related genes (TAA1, YUC3, YUC9, and NIT1) in shoots, but not in root apices. Phenotyping experiments using auxin transport-related mutants revealed that the PIN2/3/4 carriers are involved in root growth inhibition caused by B deprivation. B deprivation not only upregulated the transcriptional levels of PIN2/3/4, but also restrained the endocytosis of PIN2/3/4 carriers (observed with PIN-Dendra2 lines), resulting in elevated protein levels of PIN2/3/4 in the plasma membrane. Overall, these results suggest that B deprivation not only enhances auxin biosynthesis in shoots by elevating the expression levels of auxin biosynthesis-related genes but also promotes the polar auxin transport from shoots to roots by upregulating the gene expression levels of PIN2/3/4, as well as restraining the endocytosis of PIN2/3/4 carriers, ultimately resulting in auxin accumulation in root apices and root growth inhibition.
Assuntos
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/metabolismo , Ácidos Indolacéticos/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Boro/metabolismo , Raízes de Plantas/metabolismoRESUMO
The supply of boron (B) alleviates the toxic effects of aluminum (Al) on root growth; however, the mechanistic basis of this process remains elusive. This study filled this knowledge gap, demonstrating that boron modifies auxin distribution and transport in Al-exposed Arabidopsis roots. In B-deprived roots, treatment with Al induced an increase in auxin content in the root apical meristem zone (MZ) and transition zone (TZ), whereas in the elongation zone (EZ) the auxin content was decreased beyond the level required for adequate growth. These distribution patterns are explained by the fact that basipetal auxin transport from the TZ to the EZ was disrupted by Al-inhibited PIN-FORMED 2 (PIN2) endocytosis. Experiments involving the modulation of protein biosynthesis by cycloheximide (CHX) and transcriptional regulation by cordycepin (COR) demonstrated that the Al-induced increase of PIN2 membrane proteins was dependent upon the inhibition of PIN2 endocytosis, rather than on the transcriptional regulation of the PIN2 gene. Experiments reporting on the profiling of Al3+ and PIN2 proteins revealed that the inhibition of endocytosis of PIN2 proteins was the result of Al-induced limitation of the fluidity of the plasma membrane. The supply of B mediated the turnover of PIN2 endosomes conjugated with indole-3-acetic acid (IAA), and thus restored the Al-induced inhibition of IAA transport through the TZ to the EZ. Overall, the reported results demonstrate that boron supply mediates PIN2 endosome-based auxin transport to alleviate Al toxicity in plant roots.
Assuntos
Proteínas de Arabidopsis , Arabidopsis , Proteínas de Arabidopsis/metabolismo , Alumínio/toxicidade , Alumínio/metabolismo , Boro/metabolismo , Proteína 1 de Ligação a Repetições Teloméricas/metabolismo , Raízes de Plantas/metabolismo , Ácidos Indolacéticos/metabolismo , Arabidopsis/metabolismoRESUMO
BACKGROUND: Essential micronutrient Boron (B) plays crucial roles in plant survival and reproduction but becomes toxic in higher quantities. Although plant cells have different B transport systems, B homeostasis is mainly maintained by two transporter protein families: B exporters (BOR) and nodulin-26-like intrinsic proteins (NIP). Their diversity and differential expression are responsible for varied B tolerance among plant varieties and species. Longan is a highly admired subtropical fruit with a rising market in China and beyond. In the present study, we cultured Shixia (SX) and Yiduo (YD), two differently characterized Longan cultivars, with foliar B spray. We analyzed their leaf physiology, fruit setting, B content, and boron transporter gene expression of various tissue samples. We also traced some of these genes' subcellular localization and overexpression effects. RESULTS: YD and SX foliage share similar microstructures, except the mesophyll cell wall thickness is double in YD. The B spray differently influenced their cellular constituents and growth regulators. Gene expression analysis showed reduced BOR genes expression and NIP genes differential spatiotemporal expression. Using green fluorescent protein, two high-expressing NIPs, NIP1 and NIP19, were found to translocate in the transformed tobacco leaves' cell membrane. NIPs transformation of SX pollen was confirmed using magnetic beads and quantified using a fluorescence microscope and polymerase chain reaction. An increased seed-setting rate was observed when YD was pollinated using these pollens. Between the DlNIP1 and DlNIP19 transformed SX pollen, the former germinated better with increasing B concentrations and, compared to naturally pollinated plants, had a better seed-setting rate in YDâ × SXâ. CONCLUSION: SX and YD Longan have different cell wall structures and react differently to foliar B spray, indicating distinct B tolerance and management. Two B transporter NIP genes were traced to localize in the plasma membrane. However, under high B concentrations, their differential expression resulted in differences in Jasmonic acid content, leading to differences in germination rate. Pollination of YD using these NIPs transformed SX pollen also showed NIP1 overexpression might overcome the unilateral cross incompatibility between YDâ × SXâ and can be used to increase Longan production.
Assuntos
Boro , Proteínas de Membrana Transportadoras , Boro/metabolismo , Transporte Biológico , Proteínas de Membrana Transportadoras/genética , Plantas/metabolismo , Proteínas de Transporte/metabolismo , HomeostaseRESUMO
BACKGROUND: Boron (B) is an essential micronutrient for plants. Inappropriate B supply detrimentally affects the productivity of numerous crops. Understanding of the molecular responses of plants to different B supply levels would be of significance in crop improvement and cultivation practices to deal with the problem. RESULTS: We conducted a comprehensive analysis of the transcriptome and proteome of tobacco seedlings to investigate the expression changes of genes/proteins in response to different B supply levels, with a particular focus on B deficiency. The global gene and protein expression profiles revealed the potential mechanisms involved in the responses of tobacco to B deficiency, including up-regulation of the NIP5;1-BORs module, complex regulation of genes/proteins related to cell wall metabolism, and up-regulation of the antioxidant machinery. CONCLUSION: Our results demonstrated that B deficiency caused severe morphological and physiological disorders in tobacco seedlings, and revealed dynamic expression changes of tobacco genes/proteins in response to different B supply levels, especially to B deficiency, thus offering valuable insights into the molecular responses of tobacco to B deficiency.
Assuntos
Boro , Nicotiana , Proteoma , Transcriptoma , Boro/deficiência , Boro/metabolismo , Nicotiana/genética , Nicotiana/metabolismo , Proteoma/metabolismo , Regulação da Expressão Gênica de Plantas , Plântula/genética , Plântula/metabolismo , Plântula/crescimento & desenvolvimento , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Perfilação da Expressão GênicaRESUMO
Micronutrient application has a crucial role in mitigating salinity stress in crop plants. This study was carried out to investigate the effect of zinc (Zn) and boron (B) as foliar applications on fenugreek growth and physiology under salt stress (0 and 120 mM). After 35 days of salt treatments, three levels of zinc (0, 50, and 100 ppm) and two levels of boron (0 and 2 ppm) were applied as a foliar application. Salinity significantly reduced root length (72.7%) and shoot length (33.9%), plant height (36%), leaf area (37%), root fresh weight (48%) and shoot fresh weight (75%), root dry weight (80%) and shoot dry weight (67%), photosynthetic pigments (78%), number of branches (50%), and seeds per pod (56%). Fenugreek's growth and physiology were improved by foliar spray of zinc and boron, which increased the length of the shoot (6%) and root length (2%), fresh root weight (18%), and dry root weight (8%), and chlorophyll a (1%), chlorophyll b (25%), total soluble protein content (3%), shoot calcium (9%) and potassium (5%) contents by significantly decreasing sodium ion (11%) content. Moreover, 100 ppm of Zn and 2 ppm of B enhanced the growth and physiology of fenugreek by reducing the effect of salt stress. Overall, boron and zinc foliar spray is suggested for improvement in fenugreek growth under salinity stress.
Assuntos
Trigonella , Zinco , Boro/metabolismo , Boro/farmacologia , Clorofila A/metabolismo , Estresse Salino , Tensoativos/metabolismo , Tensoativos/farmacologia , Trigonella/metabolismo , Zinco/metabolismo , Zinco/farmacologiaRESUMO
BACKGROUND: Boron (B) is a micronutrient, but excessive levels can cause phytotoxicity, impaired growth, and reduced photosynthesis. B toxicity arises from over-fertilization, high soil B levels, or irrigation with B-rich water. Conversely, silicon (Si) is recognized as an element that mitigates stress and alleviates the toxic effects of certain nutrients. In this study, to evaluate the effect of different concentrations of Si on maize under boron stress conditions, a factorial experiment based on a randomized complete block design was conducted with three replications in a hydroponic system. The experiment utilized a nutrient solution for maize var. Merit that contained three different boron (B) concentrations (0.5, 2, and 4 mg L-1) and three Si concentrations (0, 28, and 56 mg L-1). RESULTS: Our findings unveiled that exogenous application of B resulted in a substantial escalation of B concentration in maize leaves. Furthermore, B exposure elicited a significant diminution in fresh and dry plant biomass, chlorophyll index, chlorophyll a (Chl a), chlorophyll b (Chl b), carotenoids, and membrane stability index (MSI). As the B concentration augmented, malondialdehyde (MDA) content and catalase (CAT) enzyme activity exhibited a concomitant increment. Conversely, the supplementation of Si facilitated an amelioration in plant fresh and dry weight, total carbohydrate, and total soluble protein. Moreover, the elevated activity of antioxidant enzymes culminated in a decrement in hydrogen peroxide (H2O2) and MDA content. In addition, the combined influence of Si and B had a statistically significant impact on the leaf chlorophyll index, total chlorophyll (a + b) content, Si and B accumulation levels, as well as the enzymatic activities of guaiacol peroxidase (GPX), ascorbate peroxidase (APX), and H2O2 levels. These unique findings indicated the detrimental impact of B toxicity on various physiological and biochemical attributes of maize, while highlighting the potential of Si supplementation in mitigating the deleterious effects through modulation of antioxidant machinery and biomolecule synthesis. CONCLUSIONS: This study highlights the potential of Si supplementation in alleviating the deleterious effects of B toxicity in maize. Increased Si consumption mitigated chlorophyll degradation under B toxicity, but it also caused a significant reduction in the concentrations of essential micronutrients iron (Fe), copper (Cu), and zinc (Zn). While Si supplementation shows promise in counteracting B toxicity, the observed decrease in Fe, Cu, and Zn concentrations warrants further investigation to optimize this approach and maintain overall plant nutritional status.
Assuntos
Boro , Clorofila , Hidroponia , Silício , Zea mays , Zea mays/efeitos dos fármacos , Zea mays/crescimento & desenvolvimento , Zea mays/fisiologia , Zea mays/metabolismo , Boro/toxicidade , Boro/metabolismo , Silício/farmacologia , Clorofila/metabolismo , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/crescimento & desenvolvimento , Malondialdeído/metabolismo , Carotenoides/metabolismo , Antioxidantes/metabolismo , Catalase/metabolismoRESUMO
Land plants evolved multiple adaptations to restrict transpiration. However, the underlying molecular mechanisms are not sufficiently understood. We used an ozone-sensitivity forward genetics approach to identify Arabidopsis thaliana mutants impaired in gas exchange regulation. High water loss from detached leaves and impaired decrease of leaf conductance in response to multiple stomata-closing stimuli were identified in a mutant of MURUS1 (MUR1), an enzyme required for GDP-l-fucose biosynthesis. High water loss observed in mur1 was independent from stomatal movements and instead could be linked to metabolic defects. Plants defective in import of GDP-l-Fuc into the Golgi apparatus phenocopied the high water loss of mur1 mutants, linking this phenotype to Golgi-localized fucosylation events. However, impaired fucosylation of xyloglucan, N-linked glycans, and arabinogalactan proteins did not explain the aberrant water loss of mur1 mutants. Partial reversion of mur1 water loss phenotype by borate supplementation and high water loss observed in boron uptake mutants link mur1 gas exchange phenotypes to pleiotropic consequences of l-fucose and boron deficiency, which in turn affect mechanical and morphological properties of stomatal complexes and whole-plant physiology. Our work emphasizes the impact of fucose metabolism and boron uptake on plant-water relations.
Assuntos
Arabidopsis , Fucose , Fucose/metabolismo , Guanosina Difosfato Fucose/metabolismo , Boro/metabolismo , Arabidopsis/metabolismo , Polissacarídeos/metabolismoRESUMO
Boron (B) is crucial for plant growth and development. B deficiency can impair numerous physiological and metabolic processes, particularly in root development and pollen germination, seriously impeding crop growth and yield. However, the molecular mechanism underlying boron signal perception and signal transduction is rather limited. In this study, we discovered that CPK10, a calcium-dependent protein kinase in the CPK family, has the strongest interaction with the boron transporter BOR1. Mutations in CPK10 led to growth and root development defects under B-deficiency conditions, while constitutively active CPK10 enhanced plant tolerance to B deficiency. Furthermore, we found that CPK10 interacted with and phosphorylated BOR1 at the Ser689 residue. Through various biochemical analyses and complementation of B transport in yeast and plants, we revealed that Ser689 of BOR1 is important for its transport activity. In summary, these findings highlight the significance of the CPK10-BOR1 signaling pathway in maintaining B homeostasis in plants and provide targets for the genetic improvement of crop tolerance to B-deficiency stress.
Assuntos
Proteínas de Arabidopsis , Arabidopsis , Boro , Raízes de Plantas , Boro/metabolismo , Boro/deficiência , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Fosforilação , Raízes de Plantas/metabolismo , Mutação/genética , Adaptação Fisiológica/genética , Proteínas Quinases/metabolismo , Proteínas Quinases/genética , Regulação da Expressão Gênica de Plantas , Transporte Biológico , Antiporters/metabolismo , Antiporters/genética , Ligação Proteica , Transdução de SinaisRESUMO
Soil salinity is a major environmental constraint affecting the sustainability and profitability of agricultural production systems. Salinity stress tolerance has been present in wild crop relatives but then lost, or significantly weakened, during their domestication. Given the genetic and physiological complexity of salinity tolerance traits, agronomical solutions may be a suitable alternative to crop breeding for improved salinity stress tolerance. One of them is optimizing fertilization practices to assist plants in dealing with elevated salt levels in the soil. In this review, we analyse the causal relationship between the availability of boron (an essential metalloid micronutrient) and plant's adaptive responses to salinity stress at the whole-plant, cellular, and molecular levels, and a possibility of using boron for salt stress mitigation. The topics covered include the impact of salinity and the role of boron in cell wall remodelling, plasma membrane integrity, hormonal signalling, and operation of various membrane transporters mediating plant ionic and water homeostasis. Of specific interest is the role of boron in the regulation of H+-ATPase activity whose operation is essential for the control of a broad range of voltage-gated ion channels. The complex relationship between boron availability and expression patterns and the operation of aquaporins is also discussed.
Assuntos
Boro , Salinidade , Solo , Boro/metabolismo , Solo/química , Adaptação Fisiológica/genética , Tolerância ao Sal/genética , Plantas/metabolismo , Plantas/genética , Regulação da Expressão Gênica de PlantasRESUMO
Many nucleoside triphosphate-diphosphohydrolases (NTPDases/APYRASEs, APYs) play a key role in modulating extracellular nucleotide levels. However, the Golgi-localized APYs, which help control glycosylation, have rarely been studied. Here, we identified AtAPY1, a gene encoding an NTPDase in the Golgi apparatus, which is required for cell wall integrity and plant growth under boron (B) limited availability. Loss of function in AtAPY1 hindered cell elongation and division in root tips while increasing the number of cortical cell layers, leading to swelling of the root tip and abundant root hairs under low B stress. Further, expression pattern analysis revealed that B deficiency significantly induced AtAPY1, especially in the root meristem and stele. Fluorescent-labeled AtAPY1-GFP localized to the Golgi stack. Biochemical analysis showed that AtAPY1 exhibited a preference of UDP and GDP hydrolysis activities. Consequently, the loss of function in AtAPY1 might disturb the homoeostasis of NMP-driven NDP-sugar transport, which was closely related to the synthesis of cell wall polysaccharides. Further, cell wall-composition analysis showed that pectin content increased and borate-dimerized RG-II decreased in apy1 mutants, along with a decrease in cellulose content. Eventually, altered polysaccharide characteristics presumably cause growth defects in apy1 mutants under B deficiency. Altogether, these data strongly support a novel role for AtAPY1 in mediating responses to low B availability by regulating cell wall integrity.
Assuntos
Apirase , Proteínas de Arabidopsis , Arabidopsis , Boro , Parede Celular , Complexo de Golgi , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/enzimologia , Arabidopsis/metabolismo , Parede Celular/metabolismo , Boro/metabolismo , Boro/deficiência , Complexo de Golgi/metabolismo , Proteínas de Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Apirase/metabolismo , Apirase/genética , Regulação da Expressão Gênica de Plantas , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/genética , Raízes de Plantas/metabolismo , Meristema/genética , Meristema/crescimento & desenvolvimento , Meristema/metabolismo , Pectinas/metabolismoRESUMO
A [10B]boron agent and a nuclear imaging probe for pharmacokinetic estimation form the fundamental pair in successful boron neutron capture therapy (BNCT). However, 4-[10B]borono-l-phenylalanine (BPA), used in clinical BNCT, has undesirable water solubility and tumor selectivity. Therefore, we synthesized fluorinated and α-methylated 3-borono-l-phenylalanine (3BPA) derivatives to realize improved water solubility, tumor targetability, and biodistribution. All 3BPA derivatives exhibited over 10 times higher water solubility than BPA. Treatment with α-methylated 3BPA derivatives resulted in decreased cell uptake via l-type amino acid transporter (LAT) 2 while maintaining LAT1 recognition, thereby significantly improving LAT1/LAT2 selectivity. Biodistribution studies showed that fluorinated α-methyl 3BPA derivatives exhibited reduced boron accumulation in nontarget tissues, including muscle, skin, and plasma. Consequently, these derivatives demonstrated significantly improved tumor-to-normal tissue ratios compared to 3BPA and BPA. Overall, fluorinated α-methyl 3BPA derivatives with the corresponding radiofluorinated compounds hold potential as promising agents for future BNCT/PET theranostics.
Assuntos
Terapia por Captura de Nêutron de Boro , Neoplasias , Humanos , Boro/metabolismo , Terapia por Captura de Nêutron de Boro/métodos , Distribuição Tecidual , Neoplasias/radioterapia , Neoplasias/tratamento farmacológico , Fenilalanina/química , Água , Compostos de Boro/químicaRESUMO
Boron (B) is essential for vascular plants. Rapeseed (Brassica napus) is the second leading crop source for vegetable oil worldwide, but its production is critically dependent on B supplies. BnaA3.NIP5;1 was identified as a B-efficient candidate gene in B. napus in our previous QTL fine mapping. However, the molecular mechanism through which this gene improves low-B tolerance remains elusive. Here, we report genetic variation in BnaA3.NIP5;1 gene, which encodes a boric acid channel, is a key determinant of low-B tolerance in B. napus. Transgenic lines with increased BnaA3.NIP5;1 expression exhibited improved low-B tolerance in both the seedling and maturity stages. BnaA3.NIP5;1 is preferentially polar-localized in the distal plasma membrane of lateral root cap (LRC) cells and transports B into the root tips to promote root growth under B-deficiency conditions. Further analysis revealed that a CTTTC tandem repeat in the 5'UTR of BnaA3.NIP5;1 altered the expression level of the gene, which is tightly associated with plant growth and seed yield. Field tests with natural populations and near-isogenic lines (NILs) confirmed that the varieties carried BnaA3.NIP5;1Q allele significantly improved seed yield. Taken together, our results provide novel insights into the low-B tolerance of B. napus, and the elite allele of BnaA3.NIP5;1 could serve as a direct target for breeding low-B-tolerant cultivars.
Assuntos
Aquaporinas/genética , Boro/deficiência , Brassica napus/genética , Alelos , Aquaporinas/metabolismo , Ácidos Bóricos , Boro/metabolismo , Brassica napus/crescimento & desenvolvimento , Brassica napus/metabolismo , Mapeamento Cromossômico/métodos , Análise Mutacional de DNA/métodos , Expressão Gênica/genética , Regulação da Expressão Gênica de Plantas/genética , Variação Genética/genética , Melhoramento Vegetal/métodos , Raízes de Plantas/genética , Raízes de Plantas/metabolismo , Plântula/genética , Sementes/metabolismoRESUMO
Boron (B) is essential for plant growth, but toxic in excess. In several countries, soil toxic B levels are always a severe agricultural problem in arid and semi-arid regions. Phytoremediation of excess B containing soil is still in its infancy, while high B tolerant plants with elevated protein abundance of B efflux transporter were successfully established or explored. Brassica napus (B. napus) is one of the most important oil crops. However, B efflux transporters underlying excess B tolerance in B. napus remain unknown. Here, we reported that in Brassicaceae species, B. napus had four homologous genes of Arabidopsis AtBOR4 , which were renamed BnaBOR4.1, BnaBOR4.2, BnaBOR4.3 and BnaBOR4.4. BnaBOR4.1, BnaBOR4.2 and BnaBOR4.3 showed constitutive expression and BnaBOR4.4 appeared to be a pseudogene. BnaBOR4.2 and BnaBOR4.3 were expressed in inner cell layers and BnaBOR4.1 in the outer cell layer in root tip, and all were expressed in vascular tissue in the mature zone. B efflux activity assays in yeast demonstrated that BnaBOR4.1, BnaBOR4.2 and AtBOR4 but not BnaBOR4.3 had comparable levels of B transport activity. Structure-functional analysis between BnaBOR4.3 and BnaBOR4.2 demonstrated that amino acid residue substitution at position 297 (Ala vs Pro) and 427 (Met vs Leu) is critical for the B transport activity. Mutant BnaBOR4.3M427L partially restored the B efflux activity, and both mutants BnaBOR4.3A297P and BnaBOR4.3A297P&M427L fully restored B efflux activity, indicating that the Pro297 residue is critical for their function. Further validation of BnaBOR4 was accomplished by growing transgenic Arabidopsis plants under high B conditions. Taken together, our study identified two functional B efflux genes BnaBOR4.1 and BnaBOR4.2 in B. napus, and a key amino acid residue proline 297 associated with B efflux activity. This study highlights the potential of BanBOR4 genes for B. napus cultivation under high B stress.
Assuntos
Arabidopsis , Brassica napus , Brassica napus/metabolismo , Boro/toxicidade , Boro/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Proteínas de Membrana Transportadoras/metabolismo , Aminoácidos/metabolismo , Solo , Regulação da Expressão Gênica de PlantasRESUMO
Boron (B) deficiency has been shown to inhibit root cell growth and division. However, the precise mechanism underlying B deficiency-mediated root tip growth inhibition remains unclear. In this study, we investigated the role of BnaA3.NIP5;1, a gene encoding a boric acid channel, in Brassica napus (B. napus). BnaA3.NIP5;1 is expressed in the lateral root cap and contributes to B acquisition in the root tip. Downregulation of BnaA3.NIP5;1 enhances B sensitivity in B. napus, resulting in reduced shoot biomass and impaired root tip development. Transcriptome analysis was conducted on root tips from wild-type B. napus (QY10) and BnaA3.NIP5;1 RNAi lines to assess the significance of B dynamics in meristematic cells during seedling growth. Differentially expressed genes (DEGs) were significantly enriched in plant circadian rhythm and nitrogen (N) metabolism pathways. Notably, the circadian-rhythm-related gene HY5 exhibited a similar B regulation pattern in Arabidopsis to that observed in B. napus. Furthermore, Arabidopsis mutants with disrupted circadian rhythm (hy5/cor27/toc1) displayed heightened sensitivity to low B compared to the wild type (Col-0). Consistent with expectations, B deficiency significantly disrupted N metabolism in B. napus roots, affecting nitrogen concentration, nitrate reductase enzyme activity, and glutamine synthesis. Interestingly, this disruption was exacerbated in BnaA3NIP5;1 RNAi lines. Overall, our findings highlight the critical role of B dynamics in root tip cells, impacting circadian rhythm and N metabolism, ultimately leading to retarded growth. This study provides novel insights into B regulation in root tip development and overall root growth in B. napus.
Assuntos
Boro , Brassica napus , Ritmo Circadiano , Regulação da Expressão Gênica de Plantas , Nitrogênio , Raízes de Plantas , Brassica napus/genética , Brassica napus/metabolismo , Brassica napus/crescimento & desenvolvimento , Boro/metabolismo , Boro/deficiência , Nitrogênio/metabolismo , Nitrogênio/deficiência , Raízes de Plantas/metabolismo , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/genética , Ritmo Circadiano/genética , Arabidopsis/genética , Arabidopsis/metabolismo , Arabidopsis/crescimento & desenvolvimento , Plântula/metabolismo , Plântula/crescimento & desenvolvimento , Plântula/genética , Perfilação da Expressão Gênica , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismoRESUMO
Perhalogenated closo-borates represent a new class of membrane carriers. They owe this activity to their chaotropicity, which enables the transport of hydrophilic molecules across model membranes and into living cells. The transport efficiency of this new class of cluster carriers depends on a careful balance between their affinity to membranes and cargo, which varies with chaotropicity. However, the structure-activity parameters that define chaotropic transport remain to be elucidated. Here, we have studied the modulation of chaotropic transport by decoupling the halogen composition from the boron core size. The binding affinity between perhalogenated decaborate and dodecaborate clusters carriers was quantified with different hydrophilic model cargos, namely a neutral and a cationic peptide, phalloidin and (KLAKLAK)2. The transport efficiency, membrane-lytic properties, and cellular toxicity, as obtained from different vesicle and cell assays, increased with the size and polarizability of the clusters. These results validate the chaotropic effect as the driving force behind the membrane transport propensity of boron clusters. This work advances our understanding of the structural features of boron cluster carriers and establishes the first set of rational design principles for chaotropic membrane transporters.
Assuntos
Boro , Boro/química , Boro/metabolismo , Humanos , Transporte Biológico , Compostos de Boro/química , Compostos de Boro/metabolismo , Interações Hidrofóbicas e Hidrofílicas , Boratos/química , Boratos/metabolismoRESUMO
Uptake of boron (B) in rice (Oryza sativa) is mediated by the Low silicon rice 1 (OsLsi1) channel, belonging to the NOD26-like intrinsic protein III subgroup, and the efflux transporter B transporter 1 (OsBOR1). However, it is unknown how these transporters cooperate for B uptake and how they are regulated in response to B fluctuations. Here, we examined the response of these two transporters to environmental B changes at the transcriptional and posttranslational level. OsBOR1 showed polar localization at the proximal side of both the exodermis and endodermis of mature root region, forming an efficient uptake system with OsLsi1 polarly localized at the distal side of the same cell layers. Expression of OsBOR1 and OsLsi1 was unaffected by B deficiency and excess. However, although OsLsi1 protein did not respond to high B at the protein level, OsBOR1 was degraded in response to high B within hours, which was accompanied with a significant decrease of total B uptake. The high B-induced degradation of OsBOR1 was inhibited in the presence of MG-132, a proteasome inhibitor, without disturbance of the polar localization. In contrast, neither the high B-induced degradation of OsBOR1 nor its polarity was affected by induced expression of dominant-negative mutated dynamin-related protein 1A (OsDRP1AK47A) or knockout of the mu subunit (AP2M) of adaptor protein-2 complex, suggesting that clathrin-mediated endocytosis is not involved in OsBOR1 degradation and polar localization. These results indicate that, in contrast to Arabidopsis thaliana, rice has a distinct regulatory mechanism for B uptake through clathrin-independent degradation of OsBOR1 in response to high B.
Assuntos
Boro/metabolismo , Clatrina/metabolismo , Proteínas de Membrana Transportadoras/metabolismo , Oryza/genética , Oryza/metabolismo , Raízes de Plantas/metabolismo , Biossíntese de Proteínas/efeitos dos fármacos , Transporte Biológico/efeitos dos fármacos , Transporte Biológico/genética , Clatrina/genética , Produtos Agrícolas/genética , Produtos Agrícolas/metabolismo , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Genes de Plantas , Variação Genética , Genótipo , Proteínas de Membrana Transportadoras/genética , Mutação , Raízes de Plantas/genética , Plantas Geneticamente ModificadasRESUMO
Boron modulates a wide range of plant developmental processes; however, the regulation of early fruit development by boron remains poorly defined. We report here the physiological, anatomical, metabolic, and transcriptomic impact of pre-flowering boron supply on the sweet cherry fruit set and development (S1-S5 stages). Our findings revealed that endogenous boron content increased in early growth stages (S1 and S2 stages) following preflowering boron exogenous application. Boron treatment resulted in increased fruit set (S1 and S2 stages) and mesocarp cell enlargement (S2 stage). Various sugars (e.g., fructose and glucose), alcohols (e.g., myo-inositol and maltitol), organic acids (e.g., malic acid and citric acid), amino acids (e.g., valine and serine) accumulated in response to boron application during the various developmental stages (S1-S5 stages). Transcriptomic analysis at early growth (S1 and S2 stages) identified boron-responsive genes that are mainly related to secondary metabolism, amino acid metabolism, calcium-binding, ribosome biogenesis, sugar homeostasis and especially to photosynthesis. We found various boron-induced/repressed genes, including those specifically involved in growth. Several heat shock proteins displayed distinct patterns during the initial growth in boron-exposed fruit. Gene analysis also discovered several putative candidate genes like PavPIP5K9, PavWAT1, PavMIOX, PavCAD1, PavPAL1 and PavSNRK2.7, which could facilitate the investigation of the molecular rationale underlying boron function in early fruit growth. Substantial changes in the expression of numerous transcription factors, including PavbHLH25, PavATHB.12L, and PavZAT10.1,.2 were noticed in fruits exposed to boron. The current study provides a baseline of information for understanding the metabolic processes regulated by boron during sweet cherry fruit early growth and fruit development in general.
Assuntos
Prunus avium , Frutas/genética , Frutas/metabolismo , Boro/análise , Boro/metabolismo , Perfilação da Expressão Gênica , Transcriptoma , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/metabolismoRESUMO
Oilseed rape and other crops of Brassica napus have a high demand for boron (B). Boron deficiencies result in the inhibition of root growth, and eventually premature flower abortion. Understanding the genetic mechanisms underlying flower abortion in B-limiting conditions could provide the basis to enhance B-efficiency and prevent B-deficiency-related yield losses. In this study, we assessed transcriptomic responses to B-deficiency in diverse inflorescence tissues at multiple time points of soil-grown plants that were phenotypically unaffected by B-deficiency until early flowering. Whilst transcript levels of known B transporters were higher in B-deficient samples, these remained remarkably stable as the duration of B-deficiency increased. Meanwhile, GO-term enrichment analysis indicated a growing response resembling that of a pathogen or pest attack, escalating to a huge transcriptome response in shoot heads at mid-flowering. Grouping differentially expressed genes within this tissue into MapMan functional bins indicated enrichment of genes related to wounding, jasmonic acid and WRKY transcription factors. Individual candidate genes for controlling the "flowering-without-seed-setting" phenotype from within MapMan biotic stress bins include those of the metacaspase family, which have been implicated in orchestrating programmed cell death. Overall temporal expression patterns observed here imply a dynamic response to B-deficiency, first increasing expression of B transporters before recruiting various biotic stress-related pathways to coordinate targeted cell death, likely in response to as yet unidentified B-deficiency induced damage-associated molecular patterns (DAMPs). This response indicates new pathways to target and dissect to control B-deficiency-induced flower abortion and to develop more B-efficient crops.