Identification of the HAK gene family reveals their critical response to potassium regulation during adventitious root formation in apple rootstock

Adventitious root formation is a bottleneck during vegetative proliferation.Potassium(K^(+))is an essential macronutrient for plants.K^(+)accumulation from the soil and its distribution to the different plant organs is mediated by K^(+)transporters named K^(+)transporter(KT),K^(+)uptake(KUP),or high-affinity K^(+)(HAK).This study aimed to identify members of the HAK gene family in apples and to characterize the effects of K^(+)supply on adventitious root formation and on the expression of HAK genes and the genes that putatively control auxin transport,signaling,and cell fate during adventitious root formation.In this study,34 HAK genes(MdHAKs)were identified in the apple(Malus×domestica‘Golden Delicious’)genome.A phylogenetic analysis divided MdHAKs into four clusters(Ⅰ,Ⅱ,Ⅲ,andⅣ),comprising 16,1,4,and 13 genes,respectively.The syntenic relationships revealed that 62.5%of the total MdHAK genes arise from genomic duplication events.Chromosome location,domain structure,motif analysis,and physico-chemical characteristics were subsequently investigated.Furthermore,the application of K^(+)indicated the emergence of adventitious roots at 8 d and produced more adventitious roots at 16 d than the K^(+)-free control(CK)treatment.In addition,various MdHAKs showed root-specific expression in B9 apple rootstock stem cuttings and enhanced expression during the initiation and emergence stages of adventitious root formation in response to K^(+)treatment.Additionally,K^(+)treatment enhanced the expression levels of MdPIN1,MdPIN2,and MdAUX1.Further data indicated that a higher expression of MdWOX11,MdLBD16,and MdLBD29 and of cell cycle-related genes contributed to the auxin-stimulated adventitious root formation in response to K^(+).

Adventitious Root Regeneration:Molecular Basis and Influencing Factors

Plant regeneration is a self-repair of the plant body in response to adverse conditions or damaged structures,and root regeneration allows the plant body to better adapt to its environment by supplementing the roots’structure.Previous research has shown that adventitious roots can be made to occur from scratch in two ways.Studies that simulate adventitious root regeneration through natural conditions allow the regeneration process to be broadly divided into three stages:the perception of early signals,the massive accumulation of auxin,and the transformation of cell fate.The strength of regeneration,in turn,is influenced by wounding,stress,hormones,etc.This study mainly reviews the molecular mechanisms of de novo adventitious roots and discusses how the environment,hormones,and its own development in Arabidopsis thaliana.

Effects of in vitro subculture on the physiological characteristics of adventitious root formation in microshoots of Castanea mollissima cv.'yanshanhong'

Microshoots of Castanea mollissima cv.＇yanshanhong＇ in vitro acquired an enhanced rooting capability with increasing numbers of subculture.In this study,we investigated the effect of successive subculture on adventitious root formation in vitro by the determination of the endogenous hormone level and the enzyme activity.The levels of indoleacetic acid（IAA）,abscisic acid（ABA）,cytokinins（CTK） and gibberellic acid（GA3） were determined by high performance liquid chromatography（HPLC）,and the activities of indoleacetic acid oxidase（IAAO）,peroxidase oxidase（POD）,and polyphenol oxidase（PPO） were measured by ultraviolet-spectrophotometer assay after the induction of rooting at 2nd,4th,6th and 8th subculture.The relationships between physiological characteristics and subculture numbers or rooting rate were as follows：The levels of endogenous IAA in microshoots gradually increased,and endogenous levels of ABA,CTK and GA3 in microshoots decreased slightly after serial subcultures.The level of IAA was highly correlated with subculture numbers and rooting rates.The ratios of IAA/ABA and IAA/CTK both acutely raised with increasing rooting rate during successive subcultures and had high correlations with rooting rate.The activity of IAAO and POD are significantly negatively related with subculture numbers,and the activity of PPO increased after subcultures.

Adventitious root formation in rice requires OsGNOM1 and is mediated by the OsPINs family

The fibrous root system in cereals comprises primarily adventitious roots （ARs）, which play important roles in nutrient and water uptake. Current knowledge regarding the molecular mechanism underlying AR development is still limited. We report here the isolation of four rice （Oryza sativa L.） mutants, from different genetic backgrounds, all of which were defective in AR formation. These mutants exhibited reduced numbers of lateral roots （LRs） and partial loss of gravitropism. The mutants also displayed enhanced sensitivity to N-l-naphthylphthalamic acid, an inhibitor of polar auxin transport （PAT）, indicating that the mutations affected auxin transport. Positional cloning using one of the four mutants revealed that it was caused by loss-of-function of a guanine nucleotide exchange factor for ADP- ribosylation factor （OsGNOM1）. RT-PCR and analysis of promoter：：GUS transgenic plants showed that OsGNOM1 is expressed in AR primordia, vascular tissues, LRs, root tips, leaves, anthers and lemma veins, with a distribution pattern similar to that of auxin. In addition, the expressions of OsPIN2, OsPIN5b and OsPIN9 were altered in the mutants. Taken together, these findings indicate that OsGNOM1 affects the formation of ARs through regulating PAT.

Changes in endogenous hormone levels and redox status during enhanced adventitious rooting by rare earth element neodymium of Dendrobium densiflorum shoot cuttings

The effects of neodymium nitrate （Nd^3＋） on the adventitious rooting of Dendrobium densiflorum shoot cuttings were studied. The addition of Nd^3＋ （5 μmol/L） to culture medium significantly increased rooting frequency. Histological investigation showed that Nd^3＋ did not change the process of root initiation. Nd^3＋ did not influence total endogenous cytokinin levels, but significantly increased the level of endogenous indole-3-acetic acid （IAA） in the base of shoot cuttings. Compared to the control, the ratio of IAA/cytokinins was very high in the Nd^3＋ treatment. These results suggested that the enhanced rooting frequency may be related to the increase in endogenous IAA level in Nd^3＋ treatment. Analysis of enzyme activities showed that the enhanced accumulation of the endogenous IAA by Nd^3＋ should not be attributed to inhibition of IAA decomposition by IAA oxidase or promotion of cytokinin decomposition by cytokinin oxidase. Besides, Nd^3＋ increased the ratio of reduced glutathione （GSH） and oxidized glutathione （GSSG） in the process of adventitious rooting while the ratio of ascorbate （ASC） to dehydroascorbate （DHA） was not affected.

Advances on Internal Influence Factors of Adventitious Rooting of Forest Trees

This paper reviews the internal influence factors of adventitious root, in- ducing oxidase activity, endogenous hormones, rooting inhibiting substances, and fu- ture research ideas are also proposed.

Genome-wide identification of WOX gene family in apple and a functional analysis of MdWOX4b during adventitious root formation

The plant-specific WUSCHEL-related homeobox(WOX)genes are crucial for plant growth and development.Here,we systematically identified the MdWOX gene family in apple at the genome-wide level,and analyzed the phylogenetic relationships,conserved motifs,gene structure,and syntenic relationships of the MdWOX genes.A total of 18 MdWOX genes were identified and phylogenetic analysis placed them into three clades.The phylogenetic relationships among the WOXs were further supported by the analyses of gene structure and conserved motifs.Chromosomal distribution and synteny analysis revealed that whole-genome and segmental duplications have played key roles in MdWOX gene family expansion.Moreover,the MdWOX genes exhibit tissue-specific expression patterns and MdWOX4a,MdWOX4b,MdWOX5b,MdWOX11/12a,and MdWOX11/12b may play essential roles in adventitious root development.The adventitious rooting ability was enhanced in MdWOX4b transgenic tobacco lines.The results of this study provide useful information for future functional studies on MdWOXs in the development of apple rootstocks.

Regulation of phytohormones on root primordium initiation and adventitious root formation in the etiolated shoots of Paeonia suffruticosa'Yinfen Jinlin'

Tree peony is well known and sought after for its large, colorful flowers. Its propagation is via vegetative methods. Mech- nisms of the adventitious rooting and the regulation of rooting processes are the principles and techniques of plant propagation and improvement. Microstructures and fluctuations of phytohormones in the adventitious rooting were studied with the etiolated soft- wood shoots of Paeonia suffkuticosa ＇Yinfen Jinlin＇. There are no pre-primordia in the shoots of the cultivar. Adventitious roots are produced in five stages： shoot selection, primordium initiation, primordium growth, conducting tissue differentiation and root protru- sion. Primordia initiated in the cortex. The contents of the endogenous hormones, IAA, ABA and GA, were 5.842, 0.873 and 1.043 nmol·g^-1 FW on the bases of shoots, respectively. CTKs which included isopentenyl adenine （iPA）, zeatin riboside （ZR） and dihy- drozeatin riboside （DHZR） were 0.949, 0.695 and 2.034 nmol·g^-1 FW, respectively. DHZR is active among CTKs. The ratio of IAA to GA, CTK and ABA clearly increased at the stage of primordium initiation, while they showed low levels at the stages of primor- dium growth. The ratios were restored at the shoot levels at the stage of root protrusion. IBA provoked primordia initiation in the cortex, the vascular cambium, the pith and even in the callus induced on the base of shoots. ]AA levels in the treated shoots increased gradually to its highest level （three times of control） at the stage of conducting tissue differentiation. The ratios of IAA to GA, CTK and ABA clearly decreased at the stage of primordium initiation. The ratio of IAA to ABA is regulated at 10：1.

Contribution of Adventitious Roots to Water Use Strategy of Juniperus sabina in a Semiarid Area of China

duniperus sabina Linnaeus, an evergreen shrub with prostrate life form, can effectively prevent sand moving and is an important tree species for reforestation in semiarid areas of China. It has laterally distributed adventitious roots and a deeply distributed main root system. To detect water movement between the main root system and adventitious roots, we adopted heat pulse sensors using the Heat Ratio Method, a high precision method for measuring low sap flow rates. Two sensors were implanted in each individual in the stem between the main root system and adventitious roots, and another two in lateral stems distal to all the roots. Positive sap flows during nighttime, even under saturated air moisture conditions, were detected only between the main root system and adventitious roots under drought conditions, and the rate of flow increased as drought progressed and decreased or disappeared after rain events. The results demonstrated the existence of water movement from the main root system to adventitious roots, and combined with the high contribution of nighttime sap flow to transpiration （11%-16%） the results indicate that it also involves the process of hydraulic lift, a water movement from moist subsoil to dry surface soils. Integrated water use strategy between the main root system and adventitious roots via the process of hydraulic lift of soil water maximizes water acquisition efficiency from both subsoil water and water from rain pulses on the soil surface; this increases survivability in the water-limited environment of semiarid areas.

Optimization of Factors Influencing Adventitious Rooting in Hybrid Larch

Optimization of in vitro adventitious root induction contributes to the development of a large-scale production system of hybrid clone seedlings of larch(Larix spp.).We used orthogonal testing to investigate the factors that affect the rooting of hybrid larch shoots-activated carbon,vitamin B1,sucrose,glycine,glutamic acid(Glu),cysteine,pH,and induction time in the dark and indole-3-butyric acid(IBA).Variance analysis showed that the effects of sucrose concentration on rooting rate,survival rate,and average root number,pH on the average number of roots;and Glu on survival rate were all significant(p<0.05)when compared to the mock treatments.Among all of the 45 treatments,the highest rooting rate was 90%,the highest survival rate was 100%,and the highest average root number was 11.8.The theoretically optimal procedure of in vitro root induction of hybrid larch shoots is shoot induction on L9 modified medium(pH 6.7)containing 2 mg.L^(-1)naphthalene acetic acid,0.25 mg.L^(-1)IBA,1/3 macroelements,7.5 g.L^(-1)sucrose,and 5 mg.L^(-1)Glu for 8 days,and then root induction on L9 modified medium without auxin.

Proteomic Analysis through Adventitious Rooting of Pinus radiata Stem Cuttings with Different Rooting Capabilities

In forest production systems, vegetative propagation of elite clones through adventitious rooting is a common practice. In Chile, adventitious rooting is the main methodology for vegetative reproduction of Pinus radiata. However, the capability of produce adventitious roots in gymnosperms decreases with aging. While it is true that some efforts have been made to identify markers or/and regulators of the aging process and adventitious rooting, molecular mechanisms that regulate both processes are scarcely known, especially at protein level. This research evaluated qualitative and quantitative changes in protein accumulation during the adventitious rooting process of P. radiata stem cuttings, with different rooting capabilities. Beside, an analysis of morpho-anatomical changes was performed in stem cuttings with high and low rooting capabilities, during the adventitious rooting process. It was observed that juvenile 1-year-old stem cuttings rooted in a 100%, while aged stem cuttings (3-year-old) presented only a 20% of rooting. According to the results of differential protein accumulation, univariate and multivariate analysis indicated that in total, 114 and 89 proteins were differentially accumulated in juvenile and aged cuttings, respectively. Also, identification of such proteins showed the presence of proteins related to cell wall organization and the presence of a protein related with proper distribution of auxin PIN transporter, both key in the new meristem formation process during adventitious rooting.

Additional far-red light promotes adventitious rooting of doubleroot-cutting grafted watermelon seedlings

Root regeneration is an important factor influencing the healing rate of graft union and the survival of double-root-cutting grafting.To date,little information is available on how to enhance root regeneration of rootstock in grafted watermelon(Citrullus lanatus)seedlings.In this study,the effects of different light treatments on root regeneration were determined.This revealed that addition of far-red light(Fr)could significantly expedite root formation in the rootstock.Moreover,the results of transcriptome analysis revealed that plant hormone pathway and auxinrelated genes were greatly induced by Fr,especially for auxin-response proteins(including CmIAA11,CmIAA17,and CmAUX28),Small auxinup RNA genes(including CmSAUR20 and CmSAUR50)and the auxin efflux transporter(CmPIN3).In addition,the expression of Phytochrome Interacting Factor(PIFs),such as CmPIF1,CmPIF3 and CmPIF7,was remarkably increased by Fr.These genes may act together to activate auxinrelated pathways under Fr treatment.Based on the results of HPLC-MS/MS analysis,the concentrations of different auxin-types in adventitious root were significantly influenced by Fr.Furthermore,the better growth of rootstock root displayed superior vasculature transport activity of the graft union with Fr treatment,which was determined by the acid magenta dyeing experiment.Therefore,all the results suggested that Fr could induce AR formation in rootstocks,which may be associated with the auxin accumulation by regulating the transcriptional level of auxinrelated and PIF genes.The findings of this study demonstrated a practicable way to shorten the healing period of graftings and improve the quality of grafted watermelon seedlings,which will provide a theoretical basis for the speeding development of industrialized seedlings production.

Transcriptome profiling of indole-3-butyric acid-induced adventitious root formation in softwood cuttings of walnut

Inducing adventitious root(AR)formation in mature walnut species(Juglans L.)is challenging.However,the AR formation of mature trees can be improved by rejuvenation.In rejuvenated cuttings,exogenous indole-3-butyric acid(IBA)is essential for AR formation,and the underlying mechanism is still not well understood.Therefore,we utilized transcriptome sequencing to investigate the mechanism of IBA-induced AR formation.Our results revealed that,in comparison to the control group,IBA treatment(9 mmol·L^(-1))significantly increased the endogenous indole-3-acetic acid(IAA)content,leading to an enhanced rooting rate.We performed RNA sequencing to identify differentially expressed genes(DEGs)between the IBA-treated and control(CK)groups at 1,2,3,and 5 days after cutting(DAC).The results showed that,compared to the control cuttings,there were 1539,889,785,and 984 up-regulated genes and 2791,2936,3017,and 1752 down-regulated genes,at 1,2,3,and 5 DAC,respectively.Analysis of RNA-seq data revealed that G-type ATP-binding cassette 36/37(ABCG36/37)and ATP-binding cassette subfamily D 1(ABCD1),associated with IBA transport,were down-regulated in the rejuvenation cuttings.In contrast,PIN-FORMED(PIN)and PINOID(PID),associated with auxin efflux,were up-regulated.We identified 49 auxin/indole-3-acetic acid(AUX/IAA)-encoding genes,including IAA1,IAA3,IAA5,IAA6,IAA8,IAA11,IAA12,IAA19,and IAA20,which were up-regulated at 1-5 DAC in the rejuvenated cuttings.This study highlights that the overexpression of JrWOX5/11 in poplar significantly enhance AR growth,as evidenced by increased root length,surface area,volume,and quantity.Moreover,the co-expression network analysis involving JrWOX11 and JrWOX5 in walnut cuttings elucidates complex genetic interactions,underscoring their pivotal role in the formation of AR.Our data supported the following molecular mechanism of IBA-induced adventitious root formation.Firstly,IBA is converted to free IAA in peroxisomes.Then,the highly concentrated IAA in the procambium and parenchyma cells induces WUSCHEL-related homeobox 11(WOX11)expression at two days.Finally,WOX11 acts redundantly to up-regulate WOX5,initiating the development of root primordia cells.

A Molecular Framework for the Control of Adventitious Rooting by TIR1/AFB2-Aux/IAADependent Auxin Signaling in Arabidopsis

In Arabidopsis thaliana,canonical auxin-dependent gene regulation is mediated by 23 transcription factors from the AUXIN RESPONSE FACTOR(ARF)family that interact with auxin/indole acetic acid repressors(Aux/IAAs),which themselves form co-receptor complexes with one of six TRANSPORT INHIBITOR*!/AUXIN-SIGNALLING F-BOX(TIR1/AFB)proteins.Different combinations of co-receptors drive specific sensing outputs,allowing auxin to control a myriad of processes.ARF6 and ARF8 are positive regulators of adventitious root initiation upstream of jasmonate,but the exact auxin co-receptor complexes controlling the transcriptional activity of these proteins has remained unknown.Here,using loss-of-function mutants we show that three Aux/IAA genes,IAA6,IAA9,and IAA17,act additively in the control of adventitious root(AR)initiation.These three IAA proteins interact with ARF6 and/or ARF8 and likely repress their activity in AR development.We show that TIR1 and AFB2 are positive regulators of AR formation and TIR1 plays a dual role in the control of jasmonic acid(JA)biosynthesis and conjugation,as several JA biosynthesis genes are up-regulated in the tir1-1 mutant.These results lead us to propose that in the presence of auxin,TIR1 and AFB2 form specific sensing complexes with IAA6,IAA9,and/or IAA17 to modulate JA homeostasis and control AR initiation.

Genome-wide Identification and Expression of ARF Gene Family during Adventitious Root Development in Hot Pepper(Capsicum annuum)

Auxin response factors(ARFs) are transcription factors that activate or repress the expression of primary/early auxin response genes by binding to auxin-responsive elements(Aux REs) in their promoter regions. The ARFs play important roles in diverse developmental processes.To explore the ARF gene family in hot pepper(Capsicum annuum L.), we performed a genome-wide identification and expression analysis. In this study, 19 pepper ARF genes(Ca ARFs) clustered into three phylogenetic groups(I, II, and III) were comprehensively analyzed. Conserved domain analysis showed that all Ca ARFs contained a B3 DNA-binding domain and a middle domain, but two members lacked the carboxyterminal dimerization(CTD) domain. The number of introns in Ca ARF genes ranged from 1 to 13 and the gene structure was similar among genes in the same phylogenetic group. Additionally, prediction of Ca ARFs promoter elements and putative targets for micro RNAs suggested that the regulation of Ca ARFs may occur at both transcriptional and posttranscriptional levels. Most Ca ARFs were expressed in more than one tested tissue, and most Ca ARFs were identified as being responsive to exogenous auxin. Moreover, time-course transcription profiles of Ca ARFs revealed their roles in adventitious rooting of hypocotyl cuttings from pepper seedlings. Therefore, our results will provide a foundation for better understanding the regulatory mechanisms and molecular functions of Ca ARFs in hot pepper.

Improvement of Growth and Periplocin Yield of Periploca sepium Adventitious Root Cultures by Altering Nitrogen Source Supply

Objective To increase the ultimate yield of periplocin in Periploca sepium adventitious root cultures by a two-stage culture based on nitrogen source.Methods Firstly,the effects of nitrogen source(NH4+-NO3-) at different ratios and different total initial nitrogen amounts on the accumulation of biomass and secondary metabolites in adventitious root cultures of P.sepium were investigated,and growth and production media for the two-stage culture based on the above results were established.Results The highest biomass and periplocin content were obtained in the culture medium of 15 mmol/L total nitrogen amount with NH4+-NO3-(1:2) and 30 mmol/L total nitrogen amount with nitrate as the sole nitrogen source.By adopting a fed-batch cultivation strategy,the dry weight adventitious root,periplocin content and yield were increased by 136%,108%,and 389%,respectively when compared with those of the control,reaching up to 8.13 g/L,157.15 μg/g,and 1277.63 μg/L,respectively.Furthermore,it was found that in the process of two-stage culture,the adventitious roots grew thicker significantly after they were transferred into production medium directly.Conclusion The ultimate yield of periplocin in P.sepium adventitious root cultures could be significantly increased by a two-stage culture based on nitrogen source.

A Kinase-Phosphatase-Transcription Factor Module Regulates Adventitious Root Emergence in Arabidopsis Root-Hypocotyl Junctions

Adventitious roots form from non-root tissues as part of normal development or in response to stress or wounding.The root primordia form in the source tissue,and during emergence the adventitious roots penetrate the inner cell layers and the epidermis;however,the mechanisms underlying this emergence remain largely unexplored.Here,we report that a regulatory module composed of the AP2/ERF transcription factor ABSCISIC ACID INSENSITIVE 4(ABI4),the MAP kinases MPK3 and MPK6,and the phosphatase PP2C12 plays an important role in the emergence of junction adventitious roots(J-ARs)from the root-hypocotyl junctions in Arabidopsis thaliana.ABI4 negatively regulates J-AR emergence,preventing the accumulation of reactive oxygen species and death of epidermal cells,which would otherwise facilitate J-AR emergence.Phosphorylation by MPK3/MPK6 activates ABI4 and dephosphorylation by PP2C12 inactivates ABI4.MPK3/MPK6 also directly phosphorylate and inactivate PP2C12 during J-AR emergence.We propose that this"double-check"mechanism increases the robustness of MAP kinase signaling and finely regulates the local programmed cell death required for J-AR emergence.

Identification of WRKY Transcription Factors Related to Saikosaponin Biosynthesis in Adventitious Roots of Bupleurum chinense

Objective To identify the genes of WRKY transcription factors（TFs） from roots of Bupleurum chinense and genes that potentially regulate saikosaponin（SS） biosynthesis.Methods Firstly,the subfamily cluster analysis was mainly based on Arabidopsis thaliana WRKYs for 27 putative WRKY TFs selected from previous transcriptome sequencing data.Secondly,qPCR was used to screen such genes of WRKY TFs that could be induced by NaCI and PEG6000 in adventitious roots of B.chinense.Meanwhile,saikosaponins（SSs） in treated adventitious roots were determined by HPLC.The roots were collected at 0,2,4,8,12,24,48,and 72 h after treatments,and 120 h only for PEG.Finally,the tissue-specific expression was analyzed on screened genes by qPCR.Results The 27 genes were grouped into three categories：There were nine in Group Ⅰ,15 in Group Ⅱ,and two in Group Ⅲ.Four genes of WRKYTFs,BCWRKY6,BCWRKY16,BCWRKY32,and BCWRKY35 were obviously induced by NaCI in adventitious roots of B.chinense,while only BCWRKY32 was induced by PEG.The content of SSs increased at different levels in NaCI and PEG6000 treatment.Three genes including BCWRKY6,BCWRKY32,and BCWRKY35,expressed most in roots,were similar to the accumulation pattern of SS.Conclusion The three WRKY genes,BCWRKY6,BCWRKY32,and BCWRKY35,may be involved in the biosynthesis of SS.

Co-cultured adventitious roots of Echinacea pallida and Echinacea purpurea inhibit lipopolysaccharide-induced inflammation via MAPK pathway in mouse peritoneal macrophages

Objective:In order to elucidate the biological activity of the Co-cultured adventitious roots(ARs) of Echinacea pallida and Echinacea purpurea and provide theoretical basis for its application,and the antiinflammatory activities and potential mechanisms of Co-cultured ARs were studied.Methods:The experimental materials were obtained by bioreactor co-culture technology and used in the activity research.In this study,mouse macrophages induced by lipopolysaccharide(LPS) were used as in vitro model.Different concentrations of AR extract(50-400 g/mL) were used to treat cells.The expression of pro-inflammatory cytokines was determined using enzyme linked immunosorbent assay.The inducible nitric oxide synthase and cyclooxygenase-2 expression,mitogen-activated protein kinase(MAPK) phosphorylation,and the inhibitor of nuclear factor-kappa B-a levels were determined by the Western blot analysis.Results:In the co-cultured ARs,total flavonoids and total caffeic acid were determined,and the contents of both bioactive compounds were significantly higher than those ARs from the single-species culture.Compared with the control group,the large amount of pro-inflammatory mediators was released after LPS stimulation.However,in the extract groups with different concentrations(25,50,and 100 g/mL),the production of these pro-inflammatory mediators was inhibited in a dose-dependent manner.Furthermore,the levels of phosphorylation of MAPK proteins,including p-p38, p-c-Jun N-terminal kinase,and p-extracellular regulated protein kinases were significantly(P <0.05) decreased in the extract groups,revealing that the AR extract probably involved in regulating the MAPK signaling pathway.Conclusion:Collectively,our findings suggested that the co-cultured ARs of E.pallida and E.purpurea can inhibit production of pro-inflammatory mediators in mouse peritoneal macrophages and possess the anti-inflammatory effect by regulating MAPK signaling pathways.

Study on the Influencing Factors and the Root Mechanism in Cutting Process of Prunus humilis(Bge). Sok

A procedure for cutting Prunus humilis（Bge）. Sok was comprehensively studied in this paper. It was found that the key factors involved in this procession were medium, rooting accelerator, concentration of rooting accelerator and type of shoot. The results showed that send was used as mediums; Treatment with 1 000 mg/L AST rooting powder No. 2 and semi-woody shoots were the optimal materials for cutting, and the rooting rate reached 88.1%. Anatomical study on rooting of Prunus hum#is（Bge）. Sok cutting has been carded out by the paraffin section method. The observation result shows that the adventitious root primordium of Prunus humili$（Bge）. Sok cutting belongs to the type of induced root primordium. The adventitiousroot primordium originates from the cross region of vascular cambium and pith rays.