Development of a rapid and efficient system for CR genes identification based on hairy root transformation in Brassicaceae

Many economically important crops and vegetables belonging to the cruciferous family are heavily endangered by clubroot disease caused by Plasmodiophora brassicae infection.Breeding of clubroot resistant cultivars based on mapping and cloning of resistant genes is commonly regarded as the most cost-effective and efficient way to fight against this disease.The traditional way of R gene functional validation requires stable transformation that is both time-and labor-consuming.In this study,a rapid and efficient hairy-root transgenic protocol mediated by Agrobacterium rhizogenes was developed.The transformation positive rate was over 80%in Brassica napus showed by GUS reporter gene and this transformation only took 1/6 of the time compared with stable transformation.The system was applicable to different B.napus varieties and other cruciferous crops including Brassica rapa and Brassica oleracea.In particular,two known CR genes,CRA3.7.1 and CRA8.2.4 were used respectively,as example to show that the system works well for CR gene study combined with subsequent P.brassicae infection in B.napus.Most importantly,it works both in over-expression that led to disease resistance,as well as in RNAi which led to disease susceptible phenotype.Therefore,this system can be used in batch-wise identification of CR genes,and also offered the possibility of manipulating key genes within the P.brassicae genome that could improve our knowledge on host-pathogen interaction.

Fast,simple,efficient Agrobacterium rhizogenes-mediated transformation system to non-heading Chinese cabbage with transgenic roots

Non-heading Chinese cabbage, a variety of Brassica campestris, is an important vegetable crop in the Yangtze River Basin of China. However,the immaturity of its stable transformation system and its low transformation efficiency limit gene function research on non-heading Chinese cabbage. Agrobacterium rhizogenes-mediated(ARM) transgenic technology is a rapid and effective transformation method that has not yet been established for non-heading Chinese cabbage plants. Here, we optimized conventional ARM approaches(one-step and two-step transformation methods) suitable for living non-heading Chinese cabbage plants in nonsterile environments. Transgenic roots in composite non-heading Chinese cabbage plants were identified using phenotypic detection, fluorescence observation, and PCR analysis. The transformation efficiency of a two-step method on four five-day-old non-heading Chinese cabbage seedlings(Suzhouqing, Huangmeigui, Wuyueman, and Sijiu Caixin) was 43.33%-51.09%, whereas using the stout hypocotyl resulted in a transformation efficiency of 54.88% for the 30-day-old Sijiu Caixin.The one-step method outperformed the two-step method;the transformation efficiency of different varieties was above 60%, and both methods can be used to obtain transgenic roots for functional studies within one month. Finally, optimized ARM transformation methods can easily,quickly, and effectively produce composite non-heading Chinese cabbage plants with transgenic roots, providing a reliable foundation for gene function research and non-heading Chinese cabbage genetic improvement breeding.

Fusarium pseudograminearum and F.culmorum affect the root system architecture of bread wheat

Yield losses of bread wheat due to crown rot can be more severe when drought conditions occur during the grain-filling period.Root architecture characteristics are important for soil exploration and belowground resource acquisition and are essential for adaptation to water-limited environments.Traits such as root angle,length and density have been strongly associated with acquisition efficiency and contribute to yield stability of the crop.The impact of crown rot pathogens on wheat root architecture is poorly understood.We examined differences in root angle,length and number,as well as dry root weight of the crown rot-susceptible bread wheat cultivar,Livingston inoculated with one of two crown rot pathogens Fusarium culmorum or Fusarium pseudograminearum in a transparent-sided root observation chamber.Significant adverse impacts on plant health and growth were revealed by visual discolouration of the leaf sheaths;fresh and dry shoot weight;leaf area of the oldest and the youngest fully expanded leaf and leaf number.Values of most recorded root system measurements were reduced when inoculated with either F.culmorum or F.pseudograminearum.In contrast,root angle was increased in the presence of F.culmorum but was not significantly changed by F.pseudograminearum.The development of whiteheads and grain losses in bread wheat caused by crown rot have previously been associated with blockages of the vascular systems.The method employed here was able to identify differences in the pathogen impacts on roots,which were not detected using previous systems.This research indicates that in the presence of F.culmorum and F.pseudograminearum infection,not only reductions in the size and biomass of the shoot system but also changes in the length,biomass and architecture of the root system could play an important role in yield loss.

Exogenous melatonin improves cotton yield under drought stress by enhancing root development and reducing root damage

The exogenous application of melatonin by the root drenching method is an effective way to improve crop drought resistance.However,the optimal concentration of melatonin by root drenching and the physiological mechanisms underlying melatonin-induced drought tolerance in cotton(Gossypium hirsutum L.)roots remain elusive.This study determined the optimal concentration of melatonin by root drenching and explored the protective effects of melatonin on cotton roots.The results showed that 50μmol L-1 melatonin was optimal and significantly mitigated the inhibitory effect of drought on cotton seedling growth.Exogenous melatonin promoted root development in drought-stressed cotton plants by remarkably increasing the root length,projected area,surface area,volume,diameter,and biomass.Melatonin also mitigated the drought-weakened photosynthetic capacity of cotton and regulated the endogenous hormone contents by regulating the relative expression levels of hormone-synthesis genes under drought stress.Melatonin-treated cotton seedlings maintained optimal enzymatic and non-enzymatic antioxidant capacities,and produced relatively lower levels of reactive oxygen species and malondialdehyde,thus reducing the drought stress damage to cotton roots(such as mitochondrial damage).Moreover,melatonin alleviated the yield and fiber length declines caused by drought stress.Taken together,these findings show that root drenching with exogenous melatonin increases the cotton yield by enhancing root development and reducing the root damage induced by drought stress.In summary,these results provide a foundation for the application of melatonin in the field by the root drenching method.

Effects of water and salt for groundwater-soil systems on root growth and architecture of Tamarix chinensis in the Yellow River Delta,China

To test the patterns of the root morphology and architecture indexes of Tamarix chinensis in response to water and salt changes in the two media of the groundwater and soil,three-year-old T.chinensis seedlings were chosen as the research object.Groundwater with four salinity levels was created,and three groundwater level(GL)were applied for each salinity treatment to measure the root growth and architecture indexes.In the fresh water and brackish water treatments,the topological index(TI)of the T.chinensis roots was close to 0.5,and the root architecture was close to a dichotomous branching pattern.In the saline water and saltwater treatments,the TI of the T.chinensis roots was large and close to 1.0,and the root architecture was close to a herringbone-like branching pattern.Under different GLs and salinities,the total root length was significantly greater than the internal link length,the external link length was greater than the internal link length,and the root system showed an outward expansion strategy.The treatment with fresh water and a GL of 1.5 m was the most suitable for T.chinensis root growth,while the root growth of T.chinensis was the worst in the treatment with saline water and a GL of 0.3 m.T.chinensis can adapt to the changes in soil water and salt by regulating the growth and morphological characteristics of the root system.T.chinensis can adapt to high-salt environments by reducing its root branching and to water deficiencies by expanding the distribution and absorption area of the root system.

The dorsal root ganglion as a target for neurorestoration in neuropathic pain

Neuropathic pain is a severe and chronic condition widely found in the general population.The reason for this is the extensive variety of damage or diseases that can spark this unpleasant constant feeling in patients.During the processing of pain,the dorsal root ganglia constitute an important region where dorsal root ganglion neurons play a crucial role in the transmission and propagation of sensory electrical stimulation.Furthermore,the dorsal root ganglia have recently exhibited a regenerative capacity that should not be neglected in the understanding of the development and resolution of neuropathic pain and in the elucidation of innovative therapies.Here,we will review the complex interplay between cells(satellite glial cells and inflammatory cells)and factors(cytokines,neurotrophic factors and genetic factors)that takes place within the dorsal root ganglia and accounts for the generation of the aberrant excitation of primary sensory neurons occurring in neuropathic pain.More importantly,we will summarize an updated view of the current pharmacologic and nonpharmacologic therapies targeting the dorsal root ganglia for the treatment of neuropathic pain.

Biomechanical Response of the Root System in Tomato Seedlings under Wind Disturbance

Wind disturbance as a green method can effectively prevent the overgrowth of tomato seedlings,and its mechanism may be related to root system mechanics.This study characterized the biophysical mechanical properties of taproot and lateral roots of tomato seedlings at five seedling ages and seedling substrates with three different moisture content.The corresponding root system-substrate finite element(FE)model was then developed and validated.The study showed that seedling age significantly affected the biomechanical properties of the taproot and lateral roots of the seedlings and that moisture content significantly affected the biomechanical properties of the seedling substrate(p<0.05).The established FE model was sensitive to wind speed,substrate moisture content,strong seedling index,and seedling age and was robust.The multiple linear regression equations obtained could predict the maximum stress and strain of the root system of tomato seedlings in the wind field.The strong seedling index had the greatest impact on the biomechanical response of the seedling root system during wind disturbance,followed by wind speed.In contrast,seedling age had no significant effect on the biomechanical response of the root system during wind disturbance.In the simulation,no mechanical damage was observed on the tissue of the seedling root system,but there were some strain behaviors.Based on the plant stress resistance,wind disturbance may affect the growth and development of the root system in the later growth stage.In this study,finite element and statistical analysis methods were combined to provide an effective approach for indepth analysis of the biomechanical mechanisms of wind disturbances that inhibit tomato seedlings’growth from the root system’s perspective.

Involvement of the ABA-and H_(2)O_(2)-Mediated Ascorbate-Glutathione Cycle in the Drought Stress Responses of Wheat Roots

Abscisic acid(ABA),hydrogen peroxide(H_(2)O_(2)) and ascorbate(AsA)–glutathione(GSH)cycle are widely known for their participation in various stresses.However,the relationship between ABA and H_(2)O_(2) levels and the AsA–GSH cycle under drought stress in wheat has not been studied.In this study,a hydroponic experiment was conducted in wheat seedlings subjected to 15%polyethylene glycol(PEG)6000–induced dehydration.Drought stress caused the rapid accumulation of endogenous ABA and H_(2)O_(2) and significantly decreased the number of root tips compared with the control.The application of ABA significantly increased the number of root tips,whereas the application of H_(2)O_(2) markedly reduced the number of root tips,compared with that under 15%PEG-6000.In addition,drought stress markedly increased the DHA,GSH and GSSG levels,but decreased the AsA levels,AsA/DHA and GSH/GSSG ratios compared with those in the control.The activities of the four enzymes in the AsA–GSH cycle were also markedly increased under drought stress,including glutathione reductase(GR),ascorbate peroxidase(APX),monodehydroascorbate reductase(MDHAR)and dehydroascorbate reductase(DHAR),compared with those in the control.However,the application of an ABA inhibitor significantly inhibited GR,DHAR and APX activities,whereas the application of an H_(2)O_(2) inhibitor significantly inhibited DHAR and MDHAR activities.Furthermore,the application of ABA inhibitor significantly promoted the increases of H_(2)O_(2) and the application of H_(2)O_(2) inhibitor significantly blocked the increases of ABA,compared with those under 15% PEG-6000.Taken together,the results indicated that ABA and H_(2)O_(2) probably interact under drought stress in wheat;and both of them can mediate drought stress by modulating the enzymes in AsA–GSH cycle,where ABA acts as the main regulator of GR,DHAR,and APX activities,and H_(2)O_(2) acts as the main regulator of DHAR and MDHAR activities.

Rooted Tree Optimization for Wind Turbine Optimum Control Based on Energy Storage System

The integration of wind turbines(WTs)in variable speed drive systems belongs to the main factors causing lowstability in electrical networks.Therefore,in order to avoid this issue,WTs hybridization with a storage system is a mandatory.This paper investigates WT system operating at variable speed.The system contains of a permanent magnet synchronous generator(PMSG)supported by a battery storage system(BSS).To enhance the quality of active and reactive power injected into the network,direct power control(DPC)scheme utilizing space-vector modulation(SVM)technique based on proportional-integral(PI)control is proposed.Meanwhile,to improve the rendition of this method(DPC-SVM-PI),the rooted tree optimization technique(RTO)algorithm-based controller parameter identification is used to achieve PI optimal gains.To compare the performance ofRTO-based controllers,they were implemented and tested along with some other popular controllers under different working conditions.The obtained results have shown the supremacy of the suggested PIRTO algorithm compared to competing controllers regarding total harmonic distortion(THD),overshoot percentage,settling time,rise time,average active power value,overall efficiency,and active power steadystate error.

Root Canal System Variation in Mandibular Second Molar: Middle-Mesial Canal

Middle-mesial canals in mandibular molars are present in the population depending on age, sex and ethnicity. However, limited literature alludes to its prevalence. Troughing procedures may enhance identification, cleaning and shaping. This case report expresses the recognition and management of middle-mesial canal in a mandibular second molar of 24 years old Hispanic-Latino male.

Evolution history dominantly regulates fine root lifespan in tree species across the world

Understanding the drivers of variations in fine root lifespan is key to informing nutrient cycling and productivity in terrestrial ecosystems.However,the general patterns and determinants of forest fine root lifespan at the global scale are still limited.We compiled a dataset of 421 fine root lifespan observations from 76 tree species globally to assess phylogenetic signals among species,explored relationships between fine root lifespan and biotic and abiotic factors,and quantified the relative importance of phylogeny,root system structure and functions,climatic and edaphic factors in driving global fine root lifespan variations.Overall,fine root lifespan showed a clear phylogenetic signal,with gymnosperms having a longer fine root lifespan than angiosperms.Fine root lifespan was longer for evergreens than deciduous trees.Ectomycorrhizal(ECM)plants had an extended fine root lifespan than arbuscular mycorrhizal(AM)plants.Among different climatic zones,fine root lifespan was the longest in the boreal zone,while it did not vary between the temperate and tropical zone.Fine root lifespan increased with soil depth and root order.Furthermore,the analysis of relative importance indicated that phylogeny was the strongest driver influencing the variation in forest fine root lifespan,followed by soil clay content,root order,mean annual temperature,and soil depth,while other environmental factors and root traits exerted weaker effects.Our results suggest that the global pattern of fine root lifespan in forests is shaped by the interplay of phylogeny,root traits and environmental factors.These findings necessitate accurate representations of tree evolutionary history in earth system models to predict fine root longevity and its responses to global changes.

Increasing root-lower characteristics improves drought tolerance in cotton cultivars at the seedling stage

Drought is an important abiotic stress factor in cotton production.The root system architecture(RSA)of cotton shows high plasticity which can alleviate drought-related stress under drought stress(DS)conditions;however,this alleviation is cultivar dependent.Therefore,this study estimated the genetic variability of RSA in cotton under DS.Using the paper-based growth system,we assessed the RSA variability in 80 cotton cultivars at the seedling stage,with 0 and10%polyethylene glycol 6000(PEG6000)as the control(CK)and DS treatment,respectively.An analysis of 23 aboveground and root traits in the 80 cotton cultivars revealed different responses to DS.On the 10th day after DS treatment,the degree of variation in the RSA traits under DS(5–55%)was greater than that of CK(5–49%).The 80 cultivars were divided into drought-tolerant cultivars(group 1),intermediate drought-tolerant cultivars(group 2),and drought-sensitive cultivars(group 3)based on their comprehensive evaluation values of drought resistance.Under DS,the root lengthlower,root area-lower,root volume-lower,and root length density-lower were significantly reduced by 63,71,76,and 4%in the drought-sensitive cultivars compared to CK.Notably,the drought-tolerant cultivars maintained their root lengthlower,root area-lower,root volume-lower,and root length density–lower attributes.Compared to CK,the root diameter(0–2 mm)-lower increased by 21%in group 1 but decreased by 3 and 64%in groups 2 and 3,respectively,under DS.Additionally,the drought-tolerant cultivars displayed a plastic response under DS that was characterized by an increase in the root-lower characteristics.Drought resistance was positively correlated with the root area-lower and root length density-lower.Overall,the RSA of the different cotton cultivars varied greatly under DS.Therefore,important root traits,such as the root-lower traits,provide great insights for exploring whether drought-tolerant cotton cultivars can effectively withstand adverse environments.

Root Systems Lie Algebras

A root system is any collection of vectors that has properties that satisfy the roots of a semi simple Lie algebra. If g is semi simple, then the root system A, (Q) can be described as a system of vectors in a Euclidean vector space that possesses some remarkable symmetries and completely defines the Lie algebra of g. The purpose of this paper is to show the essentiality of the root system on the Lie algebra. In addition, the paper will mention the connection between the root system and Ways chambers. In addition, we will show Dynkin diagrams, which are an integral part of the root system.

Development of an Agrobacterium-mediated CRISPR/Cas9 gene editing system in jute(Corchorus capsularis)

Jute(Corchorus capsularis L.)is the second most important natural plant fiber source after cotton.However,developing an efficient gene editing system for jute remains a challenge.In this study,the transgenic hairy root system mediated by Agrobacterium rhizogenes strain K599 was developed for Meifeng 4,an elite jute variety widely cultivated in China.The transgenic hairy root system for jute was verified by subcellular localization and bimolecular fluorescence complementation(BiFC)assays.The CHLOROPLASTOS ALTERADOS 1(CcCLA1)gene,which is involved in the development of chloroplasts,was targeted for editing at two sites in Meifeng 4.Based on this hairy root transformation,the gRNA scaffold was placed under the control of cotton ubiquitin GhU6.7 and-GhU6.9 promoters,respectively,to assess the efficiency of gene editing.Results indicated the 50.0%(GhU6.7)and 38.5%(GhU6.9)editing events in the target 2 alleles(gRNA2),but no mutation was detected in the target 1 allele(gRNA1)in transgenic-positive hairy roots.CcCLA1 gene editing at gRNA2 under the control of GhU6.7 in Meifeng 4 was also carried out by Agrobacterium tumefaciens-mediated transformation.Two CcCLA1 mutants were albinic,with a gene editing efficiency of 5.3%.These findings confirm that the CRISPR/Cas9 system,incorporating promoter GhU6.7,can be used as a gene editing tool for jute.

Shear resistance characteristics and influencing factors of root-soil composite on an alpine metal mine dump slope with different recovery periods

Artificial vegetation restoration is the main measure for vegetation restoration and soil and water conservation in alpine mine dumps on the Qinghai-Tibet Plateau,China.However,there are few reports on the dynamic changes and the influencing factors of the soil reinforcement effect of plant species after artificial vegetation restoration under different recovery periods.We selected dump areas of the Delni Copper Mine in Qinghai Province,China to study the relationship between the shear strength and the peak displacement of the root-soil composite on the slope during the recovery period,and the influence of the root traits and soil physical properties on the shear resistance characteristics of the root-soil composite via in situ direct shear tests.The results indicate that the shear strength and peak displacement of the rooted soil initially decreased and then increased with the increase of the recovery period.The shear strength of the rooted soil and the recovery period exhibited a quadratic function relationship.There is no significant function relationship between the peak displacement and the recovery period.Significant positive correlations(P<0.05)exists between the shear strength of the root-soil composite and the root biomass density,root volume density,and root area ratio,and they show significant linear correlations(P<0.05).There are no significant correlations(P>0.05)between the shear strength of the root-soil composite and the root length density,and the root volume ratio of the coarse roots to the fine roots.A significant negative linear correlation(P<0.05)exists between the peak displacement of the rooted soil and the coarse-grain content,but no significant correlations(P>0.05)with the root traits,other soil physical property indices(the moisture content and dry density of the soil),and slope gradient.The coarse-grain content is the main factor controlling the peak displacement of the rooted soil.

Hormonal and epigenetic regulation of root responses to salinity stress

Salinity stress is a major environmental stress affecting crop productivity,and its negative impact on global food security is only going to increase,due to current climate trends.Salinity tolerance was present in wild crop relatives but significantly weakened during domestication.Regaining it back requires a good understanding of molecular mechanisms and traits involved in control of plant ionic and ROS homeostasis.This review summarizes our current knowledge on the role of major plant hormones(auxin,cytokinins,abscisic acid,salicylic acid,and jasmonate)in plants adaptation to soil salinity.We firstly discuss the role of hormones in controlling root tropisms,root growth and architecture(primary root elongation,meristematic activity,lateral root development,and root hairs formation).Hormone-mediated control of uptake and sequestration of key inorganic ions(sodium,potassium,and calcium)is then discussed followed by regulation of cell redox balance and ROS signaling in salt-stressed roots.Finally,the role of epigenetic alterations such as DNA methylation and histone modifications in control of plant ion and ROS homeostasis and signaling is discussed.This data may help develop novel strategies for breeding and cultivating salt-tolerant crops and improving agricultural productivity in saline regions.

The osteoclastic activity in apical distal region of molar mesial roots affects orthodontic tooth movement and root resorption in rats

The utilization of optimal orthodontic force is crucial to prevent undesirable side effects and ensure efficient tooth movement during orthodontic treatment.However,the sensitivity of existing detection techniques is not sufficient,and the criteria for evaluating optimal force have not been yet established.Here,by employing 3D finite element analysis methodology,we found that the apical distal region(A-D region)of mesial roots is particularly sensitive to orthodontic force in rats.Tartrate-resistant acidic phosphatase(TRAP)-positive osteoclasts began accumulating in the A-D region under the force of 40 grams(g),leading to alveolar bone resorption and tooth movement.When the force reached 80 g,TRAP-positive osteoclasts started appearing on the root surface in the A-D region.Additionally,micro-computed tomography revealed a significant root resorption at 80 g.Notably,the A-D region was identified as a major contributor to whole root resorption.It was determined that 40 g is the minimum effective force for tooth movement with minimal side effects according to the analysis of tooth movement,inclination,and hyalinization.These findings suggest that the A-D region with its changes on the root surface is an important consideration and sensitive indicator when evaluating orthodontic forces for a rat model.Collectively,our investigations into this region would aid in offering valuable implications for preventing and minimizing root resorption during patients’orthodontic treatment.

Mechanism underlying the uprooting of taproot-type shrub species in the loess area of northeastern Qinghai-Xizang Plateau, China

Characteristics of root pullout resistance determine the capacity to withstand uprooting and the slope protection ability of plants.However,mechanism underlying the uprooting of taproot-type shrub species in the loess area of northeastern Qinghai-Xizang Plateau,China remains unclear.In this study,a common taproot-type shrub,Caragana korshinskii Kom.,in northeastern Qinghai-Xizang Plateau was selected as the research material.Mechanism of root-soil interaction of vertical root of C.korshinskii was investigated via a combination of a single-root pullout test and numerical simulation analysis.The results indicated that,when pulling vertically,axial force of the roots decreased with an increase in buried depth,whereas shear stress at root-soil interface initially increased and then decreased as burial depths increased.At the same buried depth,both axial force and shear stress of the roots increased with the increase in pullout force.Shear stress and plastic zone of the soil surrounding the root were symmetrically distributed along the root system.Plastic zone was located close to the surface and was caused primarily by tensile failure.In nonvertical pulling,symmetry of shear stress and plastic zone of the soil surrounding the root was disrupted.We observed larger shear stress and plastic zones on the side facing the direction of root deflection.Plastic zone included both shear and tensile failure.Axial force of the root system near the surface decreased as deflection angle of the pullout force increased.When different rainfall infiltration depths had the same vertical pulling force,root axial force decreased with the increase of rainfall infiltration depth and total root displacement increased.During rainfall infiltration,shear stress and plastic zone of the soil surrounding the root were prone to propagating deeper into the soil.These findings provide a foundation for further investigation of soil reinforcement and slope protection mechanisms of taproot-type shrub species in the loess area of northeastern Qinghai-Xizang Plateau and similar areas.

Overexpression of the peroxidase gene ZmPRX1 increases maize seedling drought tolerance by promoting root development and lignification

Drought is a main abiotic stress factor hindering plant growth,development,and crop productivity.Therefore,it is crucial to understand the mechanisms by which plants cope with drought stress.Here,the function of the maize peroxidase gene ZmPRX1 in drought stress tolerance was investigated by measurement of its expression in response to drought treatment both in a ZmPRX1 overexpression line and a mutant line.The higher root lignin accumulation and seedling survival rate of the overexpression line than that of the wild type or mutant support a role for ZmPRX1 in maize drought tolerance by regulating root development and lignification.Additionally,yeast one-hybrid,Dule luciferase and ChIP-qPCR assays showed that ZmPRX1 is negatively regulated by a nuclear-localized ZmWRKY86 transcription factor.The gene could potentially be used for breeding of drought-tolerant cultivars.

Prediction of cell-cell communication patierns of dorsal root ganglion cells:single-cell RNA sequencing data analysis

Dorsal root ganglion neurons transmit peripheral somatic information to the central nervous system,and dorsal root ganglion neuron excitability affects pain perception.Dorsal root ganglion stimulation is a new approach for managing pain sensation.Knowledge of the cell-cell communication among dorsal root ganglion cells may help in the development of new pain and itch management strategies.Here,we used the single-cell RNA-sequencing(scRNA-seq)database to investigate intercellular communication networks among dorsal root ganglion cells.We collected scRNA-seq data from six samples from three studies,yielding data on a total of 17,766 cells.Based on genetic profiles,we identified satellite glial cells,Schwann cells,neurons,vascular endothelial cells,immune cells,fibroblasts,and vascular smooth muscle cells.Further analysis revealed that eight types of dorsal root ganglion neurons mediated proprioceptive,itch,touch,mechanical,heat,and cold sensations.Moreover,we predicted several distinct forms of intercellular communication among dorsal root ganglion cells,including cell-cell contact,secreted signals,extracellular matrix,and neurotransmitter-mediated signals.The data mining predicted that Mrgpra3-positive neurons robustly express the genes encoding the adenosine Adora2b(A2B)receptor and glial cell line-derived neurotrophic factor family receptor alpha 1(GFRα-1).Our immunohistochemistry results confirmed the coexpression of the A2B receptor and GFRα-1.Intrathecal injection of the A2B receptor antagonist PSB-603 effectively prevented histamine-induced scratching behaviour in a dose-dependent manner.Our results demonstrate the involvement of the A2B receptor in the modulation of itch sensation.Furthermore,our findings provide insight into dorsal root ganglion cell-cell communication patterns and mechanisms.Our results should contribute to the development of new strategies for the regulation of dorsal root ganglion excitability.