Molecular regulatory mechanism of tooth root development

The root is crucial for the physiological function of the tooth, and a healthy root allows an artificial crown to function as required clinically. Tooth crown development has been studied intensively during the last few decades, but root development remains not well understood. Here we review the root development processes, including cell fate determination, induction of odontoblast and cementoblast differentiation, interaction of root epithelium and mesenchyme, and other molecular mechanisms. This review summarizes our current understanding of the signaling cascades and mechanisms involved in root development. It also sets the stage for de novo tooth regeneration.

Pseudomonas sp.TK35-L enhances tobacco root development and growth by inducing HRGPnt3 expression in plant lateral root formation

Rhizosphere colonization is a key requirement for the application of plant growth-promoting rhizobacteria(PGPR)as a bioferilizer.Signaling molecules are often exchanged between PGPR and plants,and genes in plants may respond to the action of PGPR.Here,the luciferase luxAB gene was electrotransformed into Pseudomonas sp.strain TK35,a PGPR with an afinity for tobacco,and the labelled TK35(TK35-L)was used to monitor colonization dynamics in the tobacco rhizosphere and evaluate the effects of colonization on tobacco growth and root development.The transcript levels of the hydroxyproline rich glycoprotein HRGPnt3 gene,a lateral root induction indicator,in tobacco roots were examined by qPCR.The results showed that TK35-L could survive for long periods in the tobacco rhizosphere and colonize new spaces in the tobacco rhizosphere following tobacco root extension,exhibiting significant increases in root development,seedling growth and potassium accumulation in tobacco plants.The upregulation of HRGPnt3 transcription in the inoculated tobacco suggested that TK35-L can promote tobacco root development by upregulating the transcript levels of the HRGPnt3 gene,which promotes tobacco seedling growth.These findings lay a foundation for future studies on the molecular mechanism underlying the plant growth-promoting activities of PGPR.Futhermore,this work provided an ideal potential strain for biofertilizer production.

Nanosilver-Promoted Lateral Root Development in Rice is Mediated through Hydrogen Peroxide

Nanosilver(10−9 m)refers to particles comprising 20–15,000 silver atoms,exhibiting high stability and specific surface area.At present,nanosilver has been used in agricultural cultivation and production.This study examined the effects of nanosilver on growth and development of rice root systems.Study results showed that fresh weight of rice belowground organs and root length both increased significantly by 5%and 25%,respectively,after rice radicles were treated with 2 ppm of nanosilver for three days.However,the H_(2)O_(2) level reached its peak at 2 days from treatment,but the activities of the antioxidant enzymes CAT,APX,and GR were inhibited by 2 ppm of nanosilver treatment.The results showed that nanosilver treatment inhibited the antioxidant enzyme activity of rice roots.The treatment of rice radicles with 5μM H_(2)O_(2) promoted root development and the same was observed when nanosilver was used for treatment.Moreover,ascorbic acid(AsA)is a H_(2)O_(2) scavenger and therefore rice root development was inhibited when AsA was added to rice radicles together with either treatment of nanosilver or H_(2)O_(2).In summary,nanosilver treatment of rice radicles promoted root growth and development via the regulation of H_(2)O_(2) and not the O2
−pathway.

The BTB/TAZ domain-containing protein CmBT1-mediated CmANR1 ubiquitination negatively regulates root development in chrysanthemum

Roots are fundamental for plants to adapt to variable environmental conditions.The development of a robust root system is orchestrated by numerous genetic determinants and,among them,the MADS-box gene ANR1 has garnered substantial attention.Prior research has demonstrated that,in chrysanthemum,CmANR1positively regulates root system development.Nevertheless,the upstream regulators involved in the CmANR1-mediated regulation of root development remain unidentified.In this study,we successfully identified bric-a-brac,tramtrack and broad(BTB)and transcription adapter putative zinc finger(TAZ)domain protein CmBT1 as the interacting partner of CmANR1 through a yeasttwo-hybrid(Y2H)screening library.Furthermore,we validated this physical interaction through bimolecular fluorescence complementation and pull-down assays.Functional assays revealed that CmBT1 exerted a negative influence on root development in chrysanthemum.In both in vitro and in vivo assays,it was evident that CmBT1mediated the ubiquitination of CmANR1 through the ubiquitin/26S proteasome pathway.This ubiquitination subsequently led to the degradation of the CmANR1 protein and a reduction in the transcription of CmANR1-targeted gene CmPIN2,which was crucial for root development in chrysanthemum.Genetic analysis suggested that CmBT1 modulated root development,at least in part,by regulating the level of CmANR1 protein.Collectively,these findings shed new light on the regulatory role of CmBT1 in degrading CmANR1 through ubiquitination,thereby repressing the expression of its targeted gene and inhibiting root development in chrysanthemum.

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.

The IDD Transcription Factors:Their Functions in Plant Development and Environmental Response

INDETERMINATE-DOMAIN proteins(IDDs)are a plant-specific transcription factor family characterized by a conserved ID domain with four zinc finger motifs.Previous studies have demonstrated that IDDs coordinate a diversity of physiological processes and functions in plant growth and development,including floral transition,plant architecture,seed and root development,and hormone signaling.In this review,we especially summarized the latest knowledge on the functions and working models of IDD members in Arabidopsis,rice,and maize,particularly focusing on their role in the regulatory network of biotic and abiotic environmental responses,such as gravity,temperature,water,and pathogens.Understanding these mechanisms underlying the function of IDD proteins in these processes is important for improving crop yields by manipulating their activity.Overall,the review offers valuable insights into the functions and mechanisms of IDD proteins in plants,providing a foundation for further research and potential applications in agriculture.

Somatic Embryogenesis Receptor Kinases Control Root Development Mainly via Brassinosteroid-Independent Actions in Arabidopsis thaliana

Brassinosteroids （BRs）, a group of plant steroidal hormones, play critical roles in many aspects of plant growth and development. Previous studies showed that BRI1-mediated BR signaling regulates cell division and differentiation during Arabidopsis root development via interplaying with auxin and other phytohormones. Arabidopsis somatic embryogenesis receptor-like kinases （SERKs）, as co-receptors of BRI1, were found to play a fundamental role in an early activation step of BR signaling pathway. Here we report a novel function of SERKs in regulating Arabidopsis root development. Genetic analyses indicated that SERKs control root growth mainly via a BR-independent pathway. Although BR signaling pathway is completely disrupted in the serkl bakl bkkl triple mutant, the root growth of the triple mutant is much severely damaged than the BR deficiency or signaling null mutants. More detailed analyses indicated that the triple mutant exhibited drastically reduced expression of a number of genes critical to polar auxin transport, cell cycle, endodermis development and root meristem differentiation, which were not observed in null BR biosynthesis mutant cpd and null BR signaling mutant bril-701.

MPK14-mediated auxin signaling controls lateral root development via ERF13-regulated very-long-chain fatty acid biosynthesis

Auxin plays a critical role in lateral root(LR)formation.The signaling module composed of auxin-response factors(ARFs)and lateral organ boundaries domain transcription factors mediates auxin signaling to control almost every stage of LR development.Here,we show that auxin-induced degradation of the APETALA2/Ethylene Responsive Factor(AP2/ERF)transcription factor ERF13,dependent on MITOGENACTIVATED PROTEIN KINASE MPK14-mediated phosphorylation,plays an essential role in LR development.Overexpression of ERF13 results in restricted passage of the LR primordia through the endodermal layer,greatly reducing LR emergence,whereas the erf13 mutants showed an increase in emerged LR.ERF13 inhibits the expression of 3-ketoacyl-CoA synthase16(KCS16),which encodes a fatty acid elongase involved in very-long-chain fatty acid(VLCFA)biosynthesis.Overexpression of KCS16 or exogenous VLCFA treatment rescues the LR emergence defects in ERF13 overexpression lines,indicating a role downstream of the auxin-MPK14-ERF13 signaling module.Collectively,our study uncovers a novel molecular mechanism by which MPK14-mediated auxin signaling modulates LR development via ERF13-regulated VLCFA biosynthesis.

Ectopic expression of foxtail millet zip-like gene,SiPf40,in transgenic rice plants causes a pleiotropic phenotype affecting tillering,vascular distribution and root development

Plant architecture determines grain production in rice(Oryza sativa) and is affected by important agronomic traits such as tillering,plant height,and panicle morphology.Many key genes involved in controlling the initiation and outgrowth of axillary buds,the elongation of stems,and the architecture of inflorescences have been isolated and analyzed.Previous studies have shown that SiPf40,which was identified from a foxtail millet(Setaria italica) immature seed cDNA library,causes extra branches and tillers in SiPf40-transgenic tobacco and foxtail millet,respectively.To reconfirm its function,we generated transgenic rice plants overexpressing SiPf40 under the control of the ubiquitin promoter.SiPf40-overexpressing transgenic plants have a greater tillering number and a wider tiller angle than wild-type plants.Their root architecture is modified by the promotion of lateral root development,and the distribution of xylem and phloem in the vascular bundle is affected.Analysis of hormone levels showed that the ratios of indole-3-acetic acid/zeatin(IAA/ZR) and IAA/gibberellic acid(IAA/GA) decreased in SiPf40-transgenic plants compared with wild-type plants.These findings strongly suggest that SiPf40 plays an important role in plant architecture.

A global alfalfa diversity panel reveals genomic selection signatures in Chinese varieties and genomic associations with root development

Alfalfa(Medicago sativa L.) is an important forage crop worldwide. However, little is known about the effects of breeding status and different geographical populations on alfalfa improvement. Here, we sequenced 220 alfalfa core germplasms and determined that Chinese alfalfa cultivars form an independent group, as evidenced by comparisons of FSTvalues between different subgroups, suggesting that geographical origin plays an important role in group differentiation. By tracing the influence of geographical regions on the genetic diversity of alfalfa varieties in China, we identified 350 common candidate genetic regions and 548 genes under selection. We also defined 165 loci associated with 24 important traits from genome-wide association studies. Of those, 17 genomic regions closely associated with a given phenotype were under selection, with the underlying haplotypes showing significant differences between subgroups of distinct geographical origins. Based on results from expression analysis and association mapping,we propose that 6-phosphogluconolactonase(MsPGL) and a gene encoding a protein with NHL domains(MsNHL) are critical candidate genes for root growth. In conclusion, our results provide valuable information for alfalfa improvement via molecular breeding.

Notes from the Underground:Receptor-Like Kinases in Arabidopsis Root Development

During plant development, the frequency and context of cell division must be controlled, and cells must differentiate properly to perform their mature functions. In addition, stem cell niches need to be maintained as a reservoir for new cells. All of these processes require intercellular signaling, whether it is a cell relaying its position to other cells, or more mature cells signaling to the stem cell niche to regulate the rate of growth. Receptor-like kinases have emerged as a major component in these diverse roles, especially within the Arabidopsis root. In this review, the functions of receptor-like kinase signaling in regulating Arabidopsis root development will be examined in theareas of root apical meristem maintenance, regulation of epidermal cell fate, lateral root development and vascular differentiation.

DNA Topoisomerase 1 Prevents R-loop Accumulation to Modulate Auxin-Regulated Root Development in Rice

R-loop structures （RNA：DNA hybrids） have important functions in many biological processes, including transcriptional regulation and genome instability among diverse organisms. DNA topoisomerase 1 （TOP1）, an essential manipulator of DNA topology during RNA transcription and DNA replication processes, can prevent R-loop accumulation by removing the positive and negative DNA supercoiling that is made by RNA polymerases during transcription. TOP1 is required for plant development, but little is known about its function in preventing co-transcriptional R-loop accumulation in various biological processes in plants. Here we show that knockdown of OsTOP1 strongly affects rice development, causing defects in root archi- tecture and gravitropism, which are the consequences of misregulation of auxin signaling and transporter genes. We found that R-loops are naturally formed at rice auxin-related gene loci, and overaccumulate when OsTOP1 is knocked down or OsTOP1 protein activity is inhibited. OsTOP1 therefore sets the accurate expression levels of auxin-related genes by preventing the overaccumulation of inherent R-loops. Our data reveal R-loops as important factors in polar auxin transport and plant root development, and highlight that OsTOP1 functions as a key to link transcriptional R-loops with plant hormone signaling, provide new in- sights into transcriptional regulation of hormone signaling in plants.

An auxin‐responsive endogenous peptide regulates root development in Arabidopsis

Auxin plays critical roles in root formation and development. The components involved in this process, however, are not well understood. Here, we newly identified a peptide encoding gene, auxin-responsive endogenous polypeptide 1 （AREP1）, which is induced by auxin, and mediates root development in Arabidopsis. Expression of AREP1 was specific to the cotyledon and to root and shoot meristem tissues. Amounts of AREP1 transcripts and AREP1-green fluorescent protein fusion proteins were elevated in response to indoleacetic acid treatment. Suppression of AREP1 through RNAi silencing resulted in reduction of primary root length, increase of lateral root number, and expansion of adventitious roots, compared to the observations in wild-type plants in the presence of auxin. By contrast, transgenic plants overexpressing AREP1 showed enhanced growth of the primary root under auxin treatment. Additionally, rootmorphology, including lateral root number and adventitious roots, differed greatly between transgenic and wildtype plants. Further analysis indicated that the expression of auxin-responsive genes, such as IAA3, IAA7, IAA17, GH3.2, GH3.3, and SAUR-AC1, was significantly higher in AREP1 RNAi plants, and was slightly lower in AREP1 overexpressing plants than in wildtype plants. These results suggest that the novel endogenous peptide AREP1 plays an important role in the process of auxinmediated root development.

OsRLR4 binds to the OsAUX1 promoter to negatively regulate primary root development in rice

Root architecture is one of the most important agronomic traits that determines rice crop yield. The primary root(PR) absorbs mineral nutrients and provides mechanical support;however, the molecular mechanisms of PR elongation remain unclear in rice. Here, the two loss-of-function T-DNA insertion mutants of root length regulator 4(Os RLR4), osrlr4-1 and osrlr4-2 with longer PR, and three Os RLR4 overexpression lines, OE-Os RLR4-1/-2/-3 with shorter PR compared to the wild type/Hwayoung(WT/HY), were identified. Os RLR4 isone of five members of the PRAF subfamily of the regulator chromosome condensation1(RCC1) family. Phylogenetic analysis of Os RLR4 from wild and cultivated rice indicated that it is under selective sweeps, suggesting its potential role in domestication. Os RLR4 controls PR development by regulating auxin accumulation in the PR tip and thus the root apical meristem activity. A series of biochemical and genetic analyses demonstrated that Os RLR4 functions directly upstream of the auxin transporter Os AUX1. Moreover, Os RLR4 interacts with the TRITHORAX-like protein Os Trx1 to promote H3 K4 me3 deposition at the Os AUX1 promoter, thus altering its transcription level. This work provides insight into the cooperation of auxin and epigenetic modifications in regulating root architecture and provides a genetic resource for plant architecture breeding.

Increase in root density induced by coronatine improves maize drought resistance in North China

Drought stress caused by insufficient irrigation or precipitation impairs agricultural production worldwide.In this study,a two-year field experiment was conducted to investigate the effect of coronatine(COR),a functional analog of jasmonic acid(JA),on maize drought resistance.The experiment included two water treatments(rainfed and irrigation),four COR concentrations(mock,0μmol L^(-1);A1,0.1μmol L^(-1);A2,1μmol L^(-1);A3,10μmol L^(-1))and two maize genotypes(Fumin 985(FM985),a drought-resistant cultivar and Xianyu 335(XY335),a drought-sensitive cultivar).Spraying 1μmol L^(-1)COR at seedling stage increased surface root density and size,including root dry matter by 12.6%,projected root area by 19.0%,average root density by 51.9%,and thus root bleeding sap by 28.2%under drought conditions.COR application also increased leaf area and SPAD values,a result attributed to improvement of the root system and increases in abscisic acid(ABA),JA,and salicylic acid(SA)contents.The improvement of leaves and roots laid the foundation for increasing plant height and dry matter accumulation.COR application reduced anthesis and silking interval,increasing kernel number per ear.COR treatment at 1μmol L^(-1)increased the yield of XY335 and FM985 by 7.9%and 11.0%,respectively.Correlation and path analysis showed that grain yields were correlated with root dry weight and projected root area,increasing maize drought resistance mainly via leaf area index and dry matter accumulation.Overall,COR increased maize drought resistance mainly by increasing root dry weight and root area,with 1μmol L-^(-1)COR as an optimal concentration.

Symplastic communication in the root cap directs auxin distribution to modulate root development

Root cap not only protects root meristem,but also detects and transduces the signals of environmental changes to affect root development.The symplastic communication is an important way for plants to transduce signals to coordinate the development and physiology in response to the changing enviroments.However,it is unclear how the symplastic communication between root cap cells affects root growth.Here we exploit an inducible system to specifically block the symplastic communication in the root cap.Transient blockage of plasmodesmata(PD)in differentiated collumella cells severely impairs the root development in Arabidopsis,in particular in the stem cell niche and the proximal meristem.The neighboring stem cell niche is the region that is most sensitive to the disrupted symplastic communication and responds rapidly via the alteration of auxin distribution.In the later stage,the cell division in proximal meristem is inhibited,presumably due to the reduced auxin level in the root cap.Our results reveal the essential role of the differentiated collumella cells in the root cap mediated signaling system that directs root development.

The hidden half comes into the spotlight: Peeking inside the black box of root developmental phases

Efficient use of natural resources(e.g.,light,water,and nutrients)can be improved with a tailored developmental program that maximizes the lifetime and fitness of plants.In plant shoots,a developmental phase represents a time window in which the meristem triggers the development of unique morphological and physiological traits,leading to the emergence of leaves,flowers,and fruits.Whereas developmental phases in plant shoots have been shown to enhance food production in crops,this phenomenon has remained poorly investigated in roots.In light of recent advances,we suggest that root development occurs in three main phases:root apical meristem appearance,foraging,and senescence.We provide compelling evidence suggesting that these phases are regulated by at least four developmental pathways:autonomous,non-autonomous,hormonal,and periodic.Root developmental pathways differentially coordinate organ plasticity,promoting morphological alterations,tissue regeneration,and cell death regulation.Furthermore,we suggest how nutritional checkpoints may allow progression through the developmental phases,thus completing the root life cycle.These insights highlight novel and exciting advances in root biology that may help maximize the productivity of crops through more sustainable agriculture and the reduced use of chemical fertilizers.

Overexpression of the LcPIN2 and LtPIN2 Gene in Arabidopsis thaliana Promotes Root Elongation

The auxin polar transporter,PIN-FORMED 2(PIN2)plays an important role in root development.However,it remains unclear whether PIN2 genes form two Liriodendron species,L.chinense(LcPIN2)and L.tulipifera(LtPIN2),are both involved in root development and whether and to what extent these two genes diverge in function.Here,we cloned and overexpressed LcPIN2 and LtPIN2 in Arabidopsis thaliana wild-type(WT)and Atpin2 mutant.Phylogenetic and sequence analysis showed a small degree of differentiation between these two Liriodendron PIN2 genes.Tissue-specific gene expression analysis indicated that both Liriodendron PIN2 genes were highly expressed in roots,implying a potential role in root development.Finally,heterologous overexpression of LcPIN2 and LtPIN2 in Arabidopsis both significantly increased the root length compared to wild-type and empty vector.Furthermore,the root length defect in Atpin2 was complemented both by LcPIN2 and LtPIN2.However,heterologous overexpression of LcPIN2 and LtPIN2 cannot rescue the defect in root gravitropism of Atpin2 mutants.Taken together,ourfindings unravel PIN2 genes from the magnoliids plant Liriodendron were functionally conserved with AtPIN2 in the dicotyledonous plant Arabidopsis in regard to the regulation of root length,but not root gravitropism.This study also provides a potential target for genetic improvement of the root system in these valuable forest trees Liriodendron.

The ABA synthesis enzyme allele OsNCED2^(T)promotes dryland adaptation in upland rice

Upland rice shows dryland adaptation in the form of a deeper and denser root system and greater drought resistance than its counterpart,irrigated rice.Our previous study revealed a difference in the frequency of the OsNCED2 gene between upland and irrigated populations.A nonsynonymous mutation(C to T,from irrigated to upland rice)may have led to functional variation fixed by artificial selection,but the exact biological function in dryland adaptation is unclear.In this study,transgenic and association analysis indicated that the domesticated fixed mutation caused functional variation in OsNCED2,increasing ABA levels,root development,and drought tolerance in upland rice under dryland conditions.OsNCED2-overexpressing rice showed increased reactive oxygen species-scavenging abilities and transcription levels of many genes functioning in stress response and development that may regulate root development and drought tolerance.OsNCED2^(T)-NILs showed a denser root system and drought resistance,promoting the yield of rice under dryland conditions.OsNCED2^(T)may confer dryland adaptation in upland rice and may find use in breeding dryland-adapted,water-saving rice.

Root Function in Nutrient Uptake and Soil Water Effect on NO3^- -N and NH4^+ -N Migration

Root function in uptake of nutrients and the effect of soil water on the transfer and distribution of NO3^--N in arable soil were studied using summer maize （Zea mays L. var. Shandan 9） as a testing crop. Results showed that root growth and water supply had a significant effect on NO3^--N transfer and made NO3^--N distributed evenly from bulk soil to rhizosphere soil. Under a natural condition with irrigation, the difference of NO3^--N concentration at different distance points from a maize plant was smaller, while obvious difference of NO3^--N concentration was observed under conditions of limited root growth space without irrigation. Whether root growth space was restricted or not, the content of soil NO3^--N decreased gradually from 10 to 0 cm from the plant, being opposite to the root absorbing area in soils. When root-grown space was limited, changes of NO3^--N concentration at different distances from a plant were similar to that of water content in tendency. Results showed that NO3^--N could be transferred as solute to plant root systems with water uptake by plants. However, the transfer and distribution of NH4^--N were not influenced by root growth and soil water supply, being different to NO3^--N.