Strigolactones modulate cotton fiber elongation and secondary cell wall thickening

Cotton is one of the most important economic crops in the world,and it is a major source of fiber in the textile industry.Strigolactones(SLs)are a class of carotenoid-derived plant hormones involved in many processes of plant growth and development,although the functions of SL in fiber development remain largely unknown.Here,we found that the endogenous SLs were significantly higher in fibers at 20 days post-anthesis(DPA).Exogenous SLs significantly increased fiber length and cell wall thickness.Furthermore,we cloned three key SL biosynthetic genes,namely GhD27,GhMAX3,and GhMAX4,which were highly expressed in fibers,and subcellular localization analyses revealed that GhD27,GhMAX3,and GhMAX4 were localized in the chloroplast.The exogenous expression of GhD27,GhMAX3,and GhMAX4 complemented the physiological phenotypes of d27,max3,and max4 mutations in Arabidopsis,respectively.Knockdown of GhD27,GhMAX3,and GhMAX4 in cotton resulted in increased numbers of axillary buds and leaves,reduced fiber length,and significantly reduced fiber thickness.These findings revealed that SLs participate in plant growth,fiber elongation,and secondary cell wall formation in cotton.These results provide new and effective genetic resources for improving cotton fiber yield and plant architecture.

Chemokine platelet factor 4 accelerates peripheral nerve regeneration by regulating Schwann cell activation and axon elongation

Schwann cells in peripheral nerves react to traumatic nerve injury by attempting to grow and regenerate.Howeve r,it is unclear what factors play a role in this process.In this study,we searched a GEO database and found that expression of platelet factor 4 was markedly up-regulated after sciatic nerve injury.Platelet factor is an important molecule in cell apoptosis,diffe rentiation,survival,and proliferation.Further,polymerase chain reaction and immunohistochemical staining confirmed the change in platelet factor 4 in the sciatic nerve at different time points after injury.Enzyme-linked immunosorbent assay confirmed that platelet factor 4 was secreted by Schwann cells.We also found that silencing platelet factor 4 decreased the proliferation and migration of primary cultured Schwann cells,while exogenously applied platelet factor 4 stimulated Schwann cell prolife ration and migration and neuronal axon growth.Furthermore,knocking out platelet factor 4 inhibited the prolife ration of Schwann cells in injured rat sciatic nerve.These findings suggest that Schwann cell-secreted platelet factor 4 may facilitate peripheral nerve repair and regeneration by regulating Schwann cell activation and axon growth.Thus,platelet factor 4 may be a potential therapeutic target for traumatic peripheral nerve injury.

Chain Elongation Using Native Soil Inocula:Exceptional n-Caproate Biosynthesis Performance and Microbial Mechanisms

This study demonstrates the feasibility and effectiveness of utilizing native soils as a resource for inocula to produce n-caproate through the chain elongation(CE)platform,offering new insights into anaerobic soil processes.The results reveal that all five of the tested soil types exhibit CE activity when supplied with high concentrations of ethanol and acetate,highlighting the suitability of soil as an ideal source for n-caproate production.Compared with anaerobic sludge and pit mud,the native soil CE system exhibited higher selectivity(60.53%),specificity(82.32%),carbon distribution(60.00%),electron transfer efficiency(165.00%),and conductivity(0.59 ms∙cm^(-1)).Kinetic analysis further confirmed the superiority of soil in terms of a shorter lag time and higher yield.A microbial community analysis indicated a positive correlation between the relative abundances of Pseudomonas,Azotobacter,and Clostridium and n-caproate production.Moreover,metagenomics analysis revealed a higher abundance of functional genes in key microbial species,providing direct insights into the pathways involved in n-caproate formation,including in situ CO_(2)utilization,ethanol oxidation,fatty acid biosynthesis(FAB),and reverse beta-oxidation(RBO).The numerous functions in FAB and RBO are primarily associated with Pseudomonas,Clostridium,Rhodococcus,Stenotrophomonas,and Geobacter,suggesting that these genera may play roles that are involved or associated with the CE process.Overall,this innovative inoculation strategy offers an efficient microbial source for n-caproate production,underscoring the importance of considering CE activity in anaerobic soil microbial ecology and holding potential for significant economic and environmental benefits through soil consortia exploration.

Candidate genes conferring ethylene-response in cultivated peanuts determined by BSA-seq and fine-mapping

Ethylene plays essential roles in plant growth,development and stress responses.The ethylene signaling pathway and molecular mechanism have been studied extensively in Arabidopsis and rice but limited in peanuts.Here,we established a sand-culture method to screen pingyangmycin mutagenized peanut lines based on their specific response to ethylene(“triple response”).An ethylene-insensitive mutant,inhibition of peanut hypocotyl elongation 1(iph1),was identified that showed reduced sensitivity to ethylene in both hypocotyl elongation and root growth.Through bulked segregant analysis sequencing,a major gene related to iph1,named AhIPH1,was preliminarily mapped at the chromosome Arahy.01,and further narrowed to a 450-kb genomic region through substitution mapping strategy.A total of 7014 genes were differentially expressed among the ACC treatment through RNA-seq analysis,of which only the Arahy.5BLU0Q gene in the candidate mapping interval was differentially expressed between WT and mutant iph1.Integrating sequence variations,functional annotation and transcriptome analysis revealed that a predicated gene,Arahy.5BLU0Q,encoding SNF1 protein kinase,may be the candidate gene for AhIPH1.This gene contained two single-nucleotide polymorphisms at promoter region and was more highly expressed in iph1 than WT.Our findings reveal a novel ethylene-responsive gene,which provides a theoretical foundation and new genetic resources for the mechanism of ethylene signaling in peanuts.

The bHLH transcription factor CsPIF4 positively regulates high temperature-induced hypocotyl elongation in cucumber

High temperature-induced hypocotyl elongation is a typical thermomorphogenesis trait that may significantly affect early seedling growth and subsequent crop yield.The ambient temperature and endogenous auxin are two critical factors that regulate hypocotyl growth.However,the mechanism of temperature and auxin integration in horticultural plants remains poorly understood.In this study,the roles of the basic helix-loop-helix transcription factor CsPIF4 in regulating auxin biosynthesis genes and the auxin content in the hypocotyl of cucumber(Cucumis sativus L.)seedlings under high temperature were investigated.qRT-PCR and in situ hybridization analysis revealed that expression of CsPIF4 was enhanced in the epidermis and vascular bundles in the hypocotyl of cucumber seedlings in response to high temperature.qRT-PCR and HPLC analysis showed that CsPIF4 positively regulated transcription of the auxin biosynthesis gene CsYUC8 and the auxin content in the hypocotyl under high temperature(35℃).The CRISPR/Cas9-mediated knockout of CsPIF4 resulted in a shorter hypocotyl compared with that of the wild type,together with decreased expression of CsYUC8 and lower auxin content in response to high temperature.Furthermore,biochemical assays showed that CsPIF4 could bind directly to the G-box motif of the CsYUC8 promoter and thereby activate CsYUC8 expression.These findings provide insight into the molecular mechanism of high temperature-mediated hypocotyl elongation in cucumber.

Evolution of pore systems in low-maturity oil shales during thermal upgrading--Quantified by dynamic SEM and machine learning

In-situ upgrading by heating is feasible for low-maturity shale oil,where the pore space dynamically evolves.We characterize this response for a heated substrate concurrently imaged by SEM.We systematically follow the evolution of pore quantity,size(length,width and cross-sectional area),orientation,shape(aspect ratio,roundness and solidity)and their anisotropy—interpreted by machine learning.Results indicate that heating generates new pores in both organic matter and inorganic minerals.However,the newly formed pores are smaller than the original pores and thus reduce average lengths and widths of the bedding-parallel pore system.Conversely,the average pore lengths and widths are increased in the bedding-perpendicular direction.Besides,heating increases the cross-sectional area of pores in low-maturity oil shales,where this growth tendency fluctuates at<300℃ but becomes steady at>300℃.In addition,the orientation and shape of the newly-formed heating-induced pores follow the habit of the original pores and follow the initial probability distributions of pore orientation and shape.Herein,limited anisotropy is detected in pore direction and shape,indicating similar modes of evolution both bedding-parallel and bedding-normal.We propose a straightforward but robust model to describe evolution of pore system in low-maturity oil shales during heating.

Lateral root elongation in maize is related to auxin synthesis and transportation mediated by N metabolism under a mixed NO_(3)^(–) and NH_(4)^(+) supply

A mixed nitrate (NO_(3)^(–)) and ammonium (NH_(4)^(+)) supply can promote root growth in maize (Zea mays),however,the changes in root morphology and the related physiological mechanism under different N forms are still unclear.Here,maize seedlings were grown hydroponically with three N supplied in three different forms (NO_(3)^(–)only,75/25 NO_(3)^(–)/NH_(4)^(+)and NH_(4)^(+)only).Compared with sole NO_(3)^(–)or NH_(4)^(+),the mixed N supply increased the total root length of maize but did not affect the number of axial roots.The main reason was the increased total lateral root length,while the average lateral root (LR) length in each axle was only slightly increased.In addition,the average LR density of 2nd whorl crown root under mixed N was also increased.Compared with sole nitrate,mixed N could improve the N metabolism of roots (such as the N influx rate,nitrate reductase (NR) and glutamine synthase (GS)enzyme activities and total amino content of the roots).Experiments with exogenously added NR and GS inhibitors suggested that the increase in the average LR length under mixed N was related to the process of N assimilation,and whether the NR mediated NO synthesis participates in this process needs further exploration.Meanwhile,an investigation of the changes in root-shoot ratio and carbon (C) concentration showed that C transportation from shoots to roots may not be the key factor in mediating lateral root elongation,and the changes in the sugar concentration in roots further proved this conclusion.Furthermore,the synthesis and transportation of auxin in axial roots may play a key role in lateral root elongation,in which the expression of ZmPIN1B and ZmPIN9 may be involved in this pathway.This study preliminarily clarified the changes in root morphology and explored the possible physiological mechanism under a mixed N supply in maize,which may provide some theoretical basis for the cultivation of crop varieties with high N efficiency.

A homeodomain-leucine zipper I transcription factor, MeHDZ14,regulates internode elongation and leaf rolling in cassava(Manihot esculenta Crantz)

Drought stress impairs plant growth and other physiological functions. MeHDZ14, a homeodomainleucine zipper I transcription factor, is strongly induced by drought stress in various cassava cultivars.However, the role of MeHDZ14 in cassava growth regulation has remained unclear. Here we report that MeHDZ14 affected plant height, such that a dwarf phenotype and altered internode elongation were observed in transgenic cassava lines. MeHDZ14 was found to negatively regulate the biosynthesis of lignin. Its overexpression resulted in abaxially rolled leaves. The morphogenesis of leaf epidermal cells was inhibited by overexpression of MeHDZ14, with decreased auxin and gibberellin and increased cytokinin contents. MeHDZ14 was found to regulate many drought-responsive genes, including genes involved in cell wall synthesis and expansion. MeHDZ14 bound to the promoter of caffeic acid 3-Omethyltransferase 1(MeCOMT1), acting as a transcriptional repressor of genes involved in cell wall development. MeHDZ14 appears to act as a negative regulator of internode elongation and epidermal cell morphogenesis during cassava leaf development.

GhIQD10 interacts with GhCaM7 to control cotton fiber elongation via calcium signaling

IQ67-domain(IQD)proteins function in plant defense and in organ development.The mechanisms by which they influence cotton fiber development are unknown.In the present study,GhIQD10 was expressed mainly in the transition period of cotton fiber development,and GhIQD10-overexpression lines showed shorter fibers.GhIQD10 interacted with GhCaM7 and the interaction was inhibited by Ca^(2+).In in vitro ovule culture,Ca^(2+)rescued the shorter-fiber phenotype of GhIQD10-overexpression lines,which were insensitive to the Ca^(2+)channel inhibitor verapamil and the Ca^(2+)pool release channel blocker 2-aminoethoxydiphenyl borate.We conclude that GhIQD10 affects cotton fiber elongation via Ca^(2+)signaling by interacting with GhCaM7.Brassinosteroid(BR)biosynthesis and signaling genes were up-regulated in GhIQD10-overexpression lines.Fiber development in these lines was not affected by epibrassinolide or the BR biosynthesis inhibitor brassinozole,indicating that the influence of GhIQD10 on fiber elongation was not associated with BR.

Analyses and identifications of quantitative trait loci and candidate genes controlling mesocotyl elongation in rice

Rice direct seeding has the significant potential to save labor and water,conserve environmental resources,and reduce greenhouse gas emissions tremendously.Therefore,rice direct seeding is becoming the major cultivation technology applied to rice production in many countries.Identifying and utilizing genes controlling mesocotyl elongation is an effective approach to accelerate breeding procedures and meet the requirements for direct-seeded rice(DSR) production.This study used a permanent mapping population with 144 recombinant inbred lines(RILs) and 2 828 bin-markers to detect quantitative trait loci(QTLs) associated with mesocotyl length in 2019 and 2020.The mesocotyl lengths of the rice RILs and their parents,Lijiangxintuanheigu(LTH) and Shennong 265(SN265),were measured in a growth chamber at 30°C in a dark environment.A total of 16 QTLs for mesocotyl length were identified on chromosomes 1(2),2(4),3(2),4,5,6,7,9,11(2),and 12.Seven of these QTLs,including qML1a,qML1b,qML2d,qML3a,qML3b,qML5,and qML11b,were reproducibly detected in both years via the interval mapping method.The major QTL,qML3a,was reidentified in two years via the composite interval mapping method.A total of 10 to 413 annotated genes for each QTL were identified in their smallest genetic intervals of 37.69 kb to 2.78 Mb,respectively.Thirteen predicted genes within a relatively small genetic interval(88.18 kb) of the major mesocotyl elongation QTL,qML3a,were more thoroughly analyzed.Finally,the coding DNA sequence variations among SN265,LTH,and Nipponbare indicated that the LOC_Os03g50550 gene was the strongest candidate gene for the qML3a QTL controlling the mesocotyl elongation.This LOC_Os03g50550 gene encodes a mitogen-activated protein kinase.Relative gene expression analysis using qRT-RCR further revealed that the expression levels of the LOC_Os03g50550 gene in the mesocotyl of LTH were significantly lower than in the mesocotyl of SN265.In conclusion,these results further strengthen our knowledge about rice’s genetic mechanisms of mesocotyl elongation.This investigation’s discoveries will help to accelerate breeding programs for new DSR variety development.

一种用于船舶汽轮机的安全且经济的启动加速概念

This paper analyses the issue of accelerated start-up of a marine steam turbine,which is an important problem because the start-up of a steam machine involves the combustion of fuel that is not transformed into useful energy.To find novel technologies that offer improvements in this aspect is essential due to restrictions on reducing ship emissions.Thus,the shorter the start-up time,the better for the environment and economy.High-pressure(HP)part of the turbine originally located on the Queen Elizabeth II unit was analysed.Advanced numerical calculations by thermal fluid-solid-interaction(Thermal FSI)were carried out.A series of simulations were performed for the accelerated start-up with controlled steam injection.A description of the chosen calculation methodology and the results obtained by simulation are included in this paper.The stress occurring during the accelerated start-up are approximately 40 MPa higher than those during the reference start-up.The relative elongations between the rotor and the hull during accelerated start-up reach a maximum value of 0.89 mm(0.83 mm for ultra-fast start-up).Reducing the steam turbine start-up time by 75%results in a 36.7 tons reduction in fuel consumption for start-up,resulting in an annual savings of 5372 USD.In conclusion,the concept proposed by the authors is safe,less expensive and does not affect the life of the turbine.In addition,results and applications from Siemens prove that additional injection of cooling steam is possible.

Fiber-specific increase of carotenoid content promotes cotton fiber elongation by increasing abscisic acid and ethylene biosynthesis

Cotton fiber is a raw material for the global textile industry and fiber quality is essential to its industrial application.Carotenoids are plant secondary metabolites that may serve as dietary components,regulate light harvesting,and scavenge reactive oxygen species.Although carotenoids accumulate predominantly in rapidly elongating cotton fibers,their roles in cotton fiber development remain poorly understood.In this study,a fiber-specific promoter proSCFP was applied to drive the expression of GhOR1Del,a positive regulator of carotenoid accumulation,to upregulate the carotenoid level in cotton fiber in planta.Fiber length,strength,and fineness were increased in proSCFP:GhOR1Del transgenic cotton and abscisic acid(ABA)and ethylene contents were increased in elongating fibers.The ABA downstream regulator GhbZIP27a stimulated the expression of the ethylene synthase gene GhACO3 by binding to its promoter,suggesting that ABA promoted fiber elongation by increasing ethylene production.These findings suggest the involvement of carotenoids and ABA signaling in promoting cotton fiber elongation and provide a strategy for improving cotton fiber quality.

ZmMs33 promotes anther elongation via modulating cell elongation regulators,metabolic homeostasis,and cell wall remodeling in maize

Plant cell elongation depends on well-defined gene regulations,adequate nutrients,and timely cell wall modifications.Anther size is positively correlated with the number and viability of pollen grains,while little is known about molecular mechanisms underlying anther cell elongation.Here,we found that properly activated cell elongation regulators at transcriptional levels in loss-of-function ZmMs33 mutant(ms33-6038)anthers failed to promote maize anther elongation.ZmMs33 deficiency disrupted metabolic homeostasis mainly by inhibiting both photosynthesis in anther endothecium and lipid accumulation in anther tapetum.Importantly,ms33-6038 anthers displayed ectopic,premature and excessive secondary cell wall thickening in anther middle layer,which constrained cell elongation structurally and blocked nutrient flows across different anther wall layers.The metabolic disorder was only found in ms33-6038 mutant rather than several representative male-sterility lines at transcriptional and post-translational levels.Collectively,the disordered metabolisms and blocked nutrient flows defeated the activated cell elongation regulators,and finally inhibited anther elongation and growth with a unique‘‘idling effect”in ms33-6038 mutant.

Investigation into a practical approach and application of cotton fiber elongation

Background The strength of cotton fiber has been extensively studied and significantly improved through selec-tive breeding,but fiber elongation has largely been ignored,even though elongation contributes to determining the energy needed to break fibers.Recent developments to calibrate the high volume instrument(HVI)for elongation has renewed interest in elongation.However,it is not understood how best to utilize yet another fiber property which has the potential to add to the complexity of fiber selection.To explore a practical approach to applying elongation,cot-ton samples were tested using single fiber methods,the Stelometer,and the HVI.Comparison of strength,elongation,and combined properties such as modulus were explored.Results HVI testing was shown to be sensitive enough to characterize elongation differences but unlike single fiber testing it was unable to capture within-sample variation.Fiber bundle testing,like Stelometer and HVI was shown to reduce bias due to fiber selection.Conclusion The use of secant modulus,an intrinsic material property,allowed for one value to represent both strength and elongation.Secant modulus was shown to contain more useful information than either elongation or work-to-break.Work-to-break was shown to be more influenced by a specific value of breaking force or elongation rather than the intrinsic behavior of the sample being tested.Exploring the influence of genetics and environment on elongation,and its interaction with other fiber properties,requires additional work.Secant modulus,by combining strength and elongation into one value,shows the potential to incorporate elongation values into fiber characteriza-tion without increasing the complexity of current fiber selection processes.

The Cotton GhWRKY91 Gene Negatively Regulates Root Elongation in Overexpressed Transgenic Arabidopsis thaliana

WRKY transcription factors play important roles in plant growth,development,and stress responses.Our previous research has shown that the GhWRKY91 gene can delay age-,abscisic acid(ABA)-,and drought-induced leaf senescence when overexpressed in transgenic Arabidopsis plants.To explore in more depth the biological functions of the GhWRKY91 gene,we further observed the root growth of overexpressing transgenic Arabidopsis thaliana under ABA and drought treatment.In this study,we transplanted the germinated seeds of wild-type(WT)and three transgenic lines(OE-12,OE-13 and OE-20)to 1/2 MS solid medium containing ABA and different concentrations of mannitol(simulated drought treatment)for culturing.The results showed that the transgenic plants had dark green leaves and short root lengths when no stress treatment was added.After ABA and mannitol treatment,the root growth of the WT and transgenic Arabidopsis was inhibited to varying degrees,and the root length downregulation of the transgenic plants was higher than that of the WT,indicating that they were more sensitive to ABA and drought.A bimolecular fluorescence complementation(BiFC)assay showed that the GhWRKY91 and GhWRKY3 proteins interact and emit yellow fluorescence in tobacco leaf cells.These results indicate that the GhWRKY91 gene negatively regulates root elongation in transgenic Arabidopsis and provide a basis for further research on the molecular mechanism of its involvement in regulating cotton root development.

Brassinosteroid Biosynthetic Gene CmDWF4 Regulates Bud Outgrowth in Chrysanthemum morifolium

Brassinosteroids(BRs),a class of steroid phytohormones,play a critical role in plant growth and development.The DWF4 gene encodes a cytochrome P450 enzyme(CYP90B1),which is considered a rate-limiting enzyme in BR biosynthesis.Here,we identified a homologous gene of DWF4 in chrysanthemum,CmDWF4.This gene was predicted to encode 491 amino acid residues with a molecular weight of 56.2 kDa and an isoelectric point(pI)of 9.10.Overexpression of CmDWF4 in chrysanthemum was found to significantly increase growth rate,number,and length of lateral buds.Transcriptome analysis showed that multiple xyloglucan endotransglycosylase/hydrolase(XTH)family encoding genes associated with cell wall modification were up-regulated in CmDWF4-overexpressing lines.qRT-PCR assay confirmed the up-regulation of CmXTH6,CmXTH23,and CmXTH28 in CmDWF4-overexpression line.Overall,this work establishes a mechanism by which BR biosynthetic gene CmDWF4 promotes lateral bud outgrowth in chrysanthemum,possibly through regulating cell elongation and expansion.

Congenital stapes suprastructure fixation presenting with fluctuating auditory symptoms:A case report

BACKGROUND Stapes ankylosis is a rare cause of conductive hearing loss,and stapes suprastructure fixation is extremely rare with fewer than 30 reported cases.Patients usually visit the clinic with non-progressive conductive hearing loss that typically began in the early years of life.CASE SUMMARY Herein,we report a case of a 37-year-old female with an isolated stapedial suprastructure fixation.The patient presented with unusual fluctuating auditory symptoms of tinnitus,ear fullness and mixed hearing loss.Pre-operative temporal bone computed tomography findings and operative findings revealed an isolated stapedial suprastructure fixation with monopod stapes caused by elongated pyramidal eminence.The hearing threshold recovered completely,and fluctuating auditory symptoms disappeared after the surgery.CONCLUSION This is the first report of stapedial suprastructure fixation with fluctuating auditory symptoms.Successful results are expected with surgical treatment.

Increased Elongation at Breaking Point with Improved Mechanical Characteristics in PLA

The main goal of this research was to increase the strength of Polylactic acid (PLA), an entirely biodegradable thermoplastic polyester, and an increase in elongation at the breaking point compared to neat PLA. To this end, S1, S2, and S3 were melt blended with various percentages of Zeolite, Glycerol, White vinegar, green camphor, Eucalyptus, and Carom seed oils. Here, the addition of glycerol, eucalyptus, and carom seed oils demonstrated an average improvement in impact and tensile strength of 13.44% and 14.55% respectively. Zeolite and glycerol work together as binding agents to improve stress transfer in the matrix, which increases tensile and flexural modulus as well as toughness elongation (>10%). The addition of the aforementioned materials led to an increase in the glass transition temperature and melting temperature, according to further DSC investigation. The thermal stability increased gradually, according to TGA data.

Eagle’s Syndrome: A Case Report of a Bilateral Elongated Styloid Process

Eagle’s syndrome is symptomatic of the styloid process (SP) elongation or calcification of the stylohyoid and stylomandibular ligaments. As a result of Eagle’s various studies, 4 out of every 100 patients showed that the styloid process was long. However only 4% of these cases are symptomatic [1]. Surgical traumas such as tonsillectomy or stylomandibular chronic irritation of the ligament, proc. of styloideusosteitis, ossifying periosteitis, tendinitis or mucositis why could it be [2]. The main symptoms are facial pain, dysphagia, sore throat, earache and headache. It may be unilateral or bilateral. The styloid process can be shortened through an intraoral or external approach. The case of Eagle syndrome in a 39-year-old female patient who was diagnosed to have bilateral elongated.

Prokaryotic Expression of Rubber Elongation Factor Gene and Preparation of Its Polyclonal Antibody

[Objective] The aim of this study was to prepare the recombination protein of rubber elongation factor and its polyclonal antibodies.[Method] The encoding gene of rubber elongation factor(REF)was amplified by RT-PCR,and cloned into the prokaryotic expression vector pDEST17 to transform into Escherichia coil BI2I-AI.The recombinant protein induced by L-Arabinose was purified by the affinity chromatography.As the immunogen,the recombination protein was used to immunize mice for preparing polyclonal antibodies,while ELISA and Western blot hybridization were used to detect the titers and specificity.[Result] The purified recombination protein of REF with high expression was used to immunize house mice for preparing polyclonal antibodies with high titer and specificity.The western blot hybridization showed that the antibody could recognize the natural REF from latex.[Conclusion] The recombination protein of REF was successfully obtained and the mouse anti REF antibody with high titer and specificity was prepared,which lays a basis for further studies on biological functions of rubber elongation factor and other membrane proteins in rubber particles.