Transcriptional regulation of MdPIN7 by MdARF19 during gravityinduced formation of adventitious root GSA in self-rooted apple stock

Self-rooted apple stock is widely used for apple production.However,the shallowness of the adventitious roots in self-rooted apple stock leads to poor performance in the barren orchards of China.This is because of the considerable difference in the development of a gravitropic set-point angle(GSA)between self-rooted apple stock and seedling rootstock.Therefore,it is crucial to study the molecular mechanism of adventitious root GSA in self-rooted apple stock for breeding self-rooted and deep-rooted apple rootstock cultivars.An apple auxin response factor MdARF19 functioned to establish the adventitious root GSA of self-rooted apple stock in response to gravity and auxin signals.MdARF19 bound directly to the MdPIN7 promoter,activating its transcriptional expression and thus regulating the formation of the adventitious root GSA in 12-2 self-rooted apple stock.However,MdARF19 influenced the expression of auxin efflux carriers(MdPIN3 and MdPIN10)and the establishment of adventitious root GSA of self-rooted apple stock in response to gravity signals by direct activation of MdFLP.Our findings provide new information on the transcriptional regulation of MdPIN7 by auxin response factor MdARF19 in the regulation of the adventitious root GSA of self-rooted apple stock in response to gravity and auxin signals.

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.

PROG1 acts upstream of LAZY1 to regulate rice tiller angle as a repressor

Rice tiller angle,as a component of plant architecture,affects rice grain yield via plant density.However,the molecular mechanism underlying rice tiller angle remains elusive.We report that the key domestication gene PROSTRATE GROWTH 1(PROG1)controls rice tiller angle by regulating shoot gravitropism and LAZY1(LA1)-mediated asymmetric distribution of auxin.Acting as a transcriptional repressor,PROG1 negatively regulates the expression of LA1 in light-grown rice seedlings.Overexpression of LA1 partially rescued the larger tiller angle of the PROG1 complementation transgenic plant(prog1-D).Double-mutant analysis showed that PROG1 acts upstream of LA1 to regulate shoot gravitropism and tiller angle.Mutation of Suppressors of lazy1(SOL1),encoding DWARF3(D3)acting in the strigolactone signal pathway,suppressed the large tiller angle of prog1-D by rescuing the transcription of LA1.The discovery of a light-sensitive PROG1-LA1 transcription regulatory module controlling rice shoot gravitropism and tiller angle sheds light on the genetic control of rice tiller angle.

A Botanist’s Cognitive View on Plant Growth: Cross-Talk between Developmental and Sensitivity Networks

An alteration in plant phenotypes assisted by their responses to the environmental stimuli (=tropism) has been fundamental to understand the “plant sensitivity ” that plays a crucial role in plants’ adaptive success. Plants succeed through the deployment of moderators controlling polar auxin-transport determining organ bending. Stimulus-specific effectors can be synthesized by the outer peripheral cells at the bending sites where they target highly conserved cellular processes and potentially persuade the plant sensitivity at large. Remarkably, the peripheral cells require different time-intervals to achieve the threshold expression-levels of stimulus-specific molecular responders. After stimulus perception, tropic curvatures (especially at growing root-apices) are duly coordinated via integrated chemical and electrical signalling which is the key to cellular communications. Thus, the acquired phenotypic alterations are the perplexed outcome of plant’s developmental pace, complemented by the sensitivity. A novel aspect of this study is to advance our understanding of plant developmental-programming and the extent of plant-sensitivity, determining the plant growth and their future applications.

LAZY1 controls rice shoot gravitropism through regulating polar auxin transport

Tiller angle of rice （Oryza sativa L.） is an important agronomic trait that contributes to grain production, and has long attracted attentions of breeders for achieving ideal plant architecture to improve grain yield. Although enormous efforts have been made over the past decades to study mutants with extremely spreading or compact tillers, the molecular mechanism underlying the control of tiller angle of cereal crops remains unknown. Here we report the cloning of the LAZY1 （LA1） gene that regulates shoot gravitropism by which the rice tiller angle is controlled. We show that LA1, a novel grass-specific gene, is temporally and spatially expressed, and plays a negative role in polar auxin transport （PAT）. Loss-of-function of LA1 enhances PAT greatly and thus alters the endogenous IAA distribution in shoots, leading to the reduced gravitropism, and therefore the tiller-spreading phenotype of rice plants.

The role of Arabidopsis 5PTase13 in root gravitropism through modulation of vesicle trafficking

Inositol polyphosphate 5-phosphatases （5PTases） are enzymes of phosphatidylinositol metabolism that affect various aspects of plant growth and development. Arabidopsis 5PTasel3 regulates auxin homeostasis and hormonerelated cotyledon vein development, and here we demonstrate that its knockout mutant 5pt13 has elevated sensitivity to gravistimulation in root gravitropic responses. The altered responses of 5pt13 mutants to 1-N-naphthylphthalamic acid （an auxin transport inhibitor） indicate that 5PTase13 might be involved in the regulation of auxin transport. Indeed, the auxin efflux carrier PIN2 is expressed more broadly under 5PTasel3 deficiency, and observations of the internalization of the membrane-selective dye FM4-64 reveal altered vesicle trafficking in 5pt13 mutants. Compared with wild-type, 5pt13 mutant seedlings are less sensitive to the inhibition by brefeldin A of vesicle cycling, seedling growth, and the intracellular cycling of the PIN1 and PIN2 proteins. Further, auxin redistribution upon gravitropic stimulation is stimulated under 5PTasel3 deficiency. These results suggest that 5PTasel3 may modulate auxin transport by regulating vesicle trafficking and thereby play a role in root gravitropism.

Auxin distribution and transport during embryogenesis and seed germi-nation of Arabidopsis

Auxin distribution during embryogenesis and seed germination were studied with transgenic Arabidopsis plants expressing GUS gene driven by a synthetic DR5 promoter,an auxin responsive promoter. The results showed that GUS activity is higher in ends of hypophysis and cotyledon primordia of heart-, torpedo- and cotyledon-stage embryos, leaf tip area, lateral root primordia, root apex and cotyledon of young seedlings. And GUS accumulated in root apex of the seedlings grown on auxin transport inhibitor containing media. All these suggested that above-mentioned part of the organs and tissues have a higher level of auxin, and auxin polar transport inhibitor could cause the accumulation of auxin in root apex. And auxin transport inhibitor also resulted in aberration of Arabidopsis leaf pattern formation, root gravitropism and elongation.

Brassinosteroid-regulated plant growth and development and gene expression in soybean

Brassinosteroids(BRs) are endogenous phytohormones that play important roles in regulating plant growth and development.In this study, we evaluated the effects of brassinolide(BL, one of the active BRs) on soybean and identified roles of the hormone in regulating multiple aspects of plant growth and development.BL application promoted hypocotyl and epicotyl elongation in the light but blocked epicotyl elongation in the dark.High levels of castasterone and BL accumulated in light-grown plants.BL disrupted shoot negative gravitropism, whereas gibberellin did not.BL delayed leaf senescence.Transcriptome analysis showed that BL induced cell wall-modifying genes and auxin-associated genes but suppressed a class of WRKY genes involved in senescence and stress responses,showing the complex roles of BRs in multiple biological processes.

Identification of a Gravitropism-Deficient Mutant in Rice

A gravitropism-deficient mutant M96 was isolated from a mutant bank, generated by ethyl methane sulfonate(EMS) mutagenesis of indica rice accession ZJ100. The mutant was characterized as prostrate growth at the beginning of germination, and the prostrate growth phenotype ran through the whole life duration. Tiller angle and tiller number of M96 increased significantly in comparison with the wild type. Tissue section observation analysis indicated that asymmetric stem growth around the second node occurred in M96. Genetic analysis and gene mapping showed that M96 was controlled by a single recessive nuclear gene, tentatively termed as gravitropism-deficient M96(gd M96), which was mapped to a region of 506 kb flanked by markers RM5960 and In Del8 on the long arm of chromosome 11. Sequencing analysis of the open reading frames in this region revealed a nucleotide substitution from G to T in the third exon of LOC_Os11g29840. Additionally, real-time fluorescence quantitative PCR analysis showed that the expression level of LOC_Os11g29840 in the stems was much higher than in the roots and leaves in M96. Furthermore, the expression level was more than four times in M96 stem than in the wild type stem. Our results suggested that the mutant gene was likely a new allele to the reported gene LAZY1. Isolation of this new allele would facilitate the further characterization of LAZY1.

The chloroplast-localized protein LTA1 regulates tiller angle and yield of rice

Plant architecture strongly influences rice grain yield.We report the cloning and characterization of the LTA1 gene,which simultaneously controls tiller angle and yield of rice.LTA1 encodes a chloroplastlocalized protein with a conserved YbaB DNA-binding domain,and is highly expressed in photosynthetic tissues including leaves and leaf sheaths.Disrupting the function of LTA1 leads to large tiller angle and yield reduction of rice.LTA1 affects the gravity response by mediating the distribution of endogenous auxin,thereby regulating the tiller angle.An lta1 mutant showed abnormal chloroplast development and decreased chlorophyll content and photosynthetic rate,in turn leading to reduction of rice yield.Our findings shed light on the genetic basis of tiller angle and provide a potential gene resource for the improvement of plant architecture and rice yield.

Preliminary study on a gravity－insensitive rice mutant

A gravity-insensitive mutant was isolated from rice (Oryza sativa L. cv. Zhonghua 11) transformed by Agrobacterium tumefaciens. The mutant's shoot growth (prostrate growth) was insensitive to gravity; whereas root growth displayed a normal positive gravitropism. Histological observation of root caps and leaf sheaths indicated that there was no significant difference in the number and size of amyloplasts in cells of the mutant and cells of the wild type.

Characterization of wavy root 1,an agravitropism allele,reveals the functions of OsPIN2 in fine regulation of auxin transport and distribution and in ABA biosynthesis and response in rice(Oryza sativa L.)

Root system architecture is influenced by gravity.How the root senses gravity and directs its orientation,so-called gravitropism,is not only a fundamental question in plant biology but also theoretically important for genetic improvement of crop root architecture.However,the mechanism has not been elucidated in most crops.We characterized a rice agravitropism allele,wavy root 1(war1),a loss-of-function allele in OsPIN2,which encodes an auxin efflux transporter.With loss of OsPIN2 function,war1 leads to altered root system architecture including wavy root,larger root distribution angle,and shallower root system due to the loss of gravitropic perception in root tips.In the war1 mutant,polar auxin transport was disrupted in the root tip,leading to abnormal auxin levels and disturbed auxin transport and distribution in columella cells.Amyloplast sedimentation,an important process in gravitropic sensing,was also decreased in root tip columella cells.The results indicated that OsPIN2 controls gravitropism by finely regulating auxin transport,distribution and levels,and amyloplast sedimentation in root tips.We identified a novel role of OsPIN2 in regulating ABA biosynthesis and response pathways.Loss of OsPIN2 function in the war1 resulted in increased sensitivity to ABA in seed germination,increased ABA level,changes in ABA-associated genes in roots,and decreased drought tolerance in the seedlings.These results suggest that the auxin transporter OsPIN2 not only modulates auxin transport to control root gravitropism,but also functions in ABA signaling to affect seed germination and root development,probably by mediating crosstalk between auxin and ABA pathways.

Arabidopsis Phytochrome D Is Involved in Red Light-Induced Negative Gravitropism of Hypocotyles

The phytochrome gene family, which is in Arabidopsis thaliana, consists of phytochromes A-E（phyA to phyE）, regulates plant responses to ambient light environments. PhyA and phyB have been characterized in detail, but studies on phyC to phyE have reported discrepant functions. In this study, we show that phyD regulates the Arabidopsis gravitropic response by inhibiting negative gravitropism of hypocotyls under red light condition. PhyD had only a limited effect on the gravitropic response of roots in red light condition. PhyD also enhanced phyB-regulated gravitropic responses in hypocotyls. Moreover, the regulation of hypocotyl gravitropic responses by phyD was dependent upon the red light fluence rate.

Light-stabilized GIL1 suppresses PIN3 activity to inhibit hypocotyl gravitropism

Light and gravity coordinately regulate the directional growth of plants.Arabidopsis Gravitropic in the Light 1(GIL1)inhibits the negative gravitropism of hypocotyls in red and far-red light,but the underlying molecular mechanisms remain elusive.Our study found that GIL1 is a plasma membrane-localized protein.In endodermal cells of the upper part of hypocotyls,GIL1 controls the negative gravitropism of hypocotyls.GIL1 directly interacts with PIN3 and inhibits the auxin transport activity of PIN3.Mutation of PIN3 suppresses the abnormal gravitropic response of gil1 mutant.The GIL1 protein is unstable in darkness but it is stabilized by red and far-red light.Together,our data suggest that light-stabilized GIL1 inhibits the negative gravitropism of hypocotyls by suppressing the activity of the auxin transporter PIN3,thereby enhancing the emergence of young seedlings from the soil.

LAZY4 acts additively with the starch-statolithdependent gravity-sensing pathway to regulate shoot gravitropism and tiller angle in rice

Rice tiller angle is a key agronomic trait that has significant effects on the establishment of a high-yield rice population.However,the molecular mechanism underlying the control of rice tiller angle remains to be clarified.Here,we characterized the novel tiller-angle gene LAZY4(LA4)in rice through map-based cloning.LA4 encodes a C3H2C3-type RING zinc-finger E3 ligase localized in the nucleus,and an in vitro ubiquitination assay revealed that the conserved RING finger domain is essential for its E3 ligase activity.We found that expression of LA4 can be induced by gravistimulation and that loss of LA4 function leads to defective shoot gravitropism caused by impaired asymmetric auxin redistribution upon gravistimulation.Genetic analysis demonstrated that LA4 acts in a distinct pathway from the starch biosynthesis regulators LA2 and LA3,which function in the starch-statolith-dependent pathway.Further genetic analysis showed that LA4 regulates shoot gravitropism and tiller angle by acting upstream of LA1 to mediate lateral auxin transport upon gravistimulation.Our studies reveal that LA4 regulates shoot gravitropism and tiller angle upstream of LA1 through a novel pathway independent of the LA2-LA3-mediated gravity-sensing mechanism,providing new insights into the rice tiller-angle regulatory network.

Single-cell network analysis reveals gene expression programs for Arabidopsis root development and metabolism

Single-cell RNA-sequencing datasets of Arabidopsis roots have been generated,but related comprehensive gene co-expression network analyses are lacking.We conducted a single-cell gene co-expression network analysis with publicly available scRNA-seq datasets of Arabidopsis roots using a SingleCellGGM algorithm.The analysis identified 149 gene co-expression modules,which we considered to be gene expression programs(GEPs).By examining their spatiotemporal expression,we identified GEPs specifically expressed in major root cell types along their developmental trajectories.These GEPs define gene programs regulating root cell development at different stages and are enriched with relevant developmental regulators.As examples,a GEP specific for the quiescent center(QC)contains 20 genes regulating QC and stem cell niche homeostasis,and four GEPs are expressed in sieve elements(SEs)from early to late developmental stages,with the early-stage GEP containing 17 known SE developmental regulators.We also identified GEPs for metabolic pathways with cell-type-specific expression,suggesting the existence of cell-type-specific metabolism in roots.Using the GEPs,we discovered and verified a columellaspecific gene,NRL27,as a regulator of the auxin-related root gravitropism response.Our analysis thus systematically reveals GEPs that regulate Arabidopsis root development and metabolism and provides ample resources for root biology studies.

The AFB1 auxin receptor controls the cytoplasmic auxin response pathway in Arabidopsis thaliana

The phytohormone auxin triggers root growth inhibition within seconds via a non-transcriptionalpathway.Among members of the TIR1/AFB auxin receptor family,AFB1 has a primary role in this rapidresponse. However, the unique features that confer this specific function have not been identified.Here we show that the N-terminal region of AFB1, including the F-box domain and residues thatcontribute to auxin binding,is essential and sufficient for its specific role in the rapid response. Substitutionof the N-terminal region of AFB1 with that of TIR1 disrupts its distinct cytoplasm-enriched localizationand activity in rapid root growth inhibition by auxin. Importantly, the N-terminal region of AFB1 isindispensable for auxin-triggered calcium influx, which is a prerequisite for rapid root growth inhibition.Furthermore, AFB1 negatively regulates lateral root formation and transcription of auxin-induced genes,suggesting that it plays an inhibitory role in canonical auxin signaling. These results suggest that AFB1may buffer the transcriptional auxin response, whereas it regulates rapid changes in cell growth thatcontributeto rootgravitropism.

Brassinosteroids Regulate Root Growth, Development, and Symbiosis

Brassinosteroids （BRs） are natural plant hormones critical for growth and development. BR-deficient or signaling mutants show significantly shortened root phenotypes. But for a long time, it was thought that these phenotypes were solely caused by reduced root cell elongation in the mutants. Functions of BRs in regulating root development have been largely neglected. Recent detailed analyses, however, revealed that BRs are not only involved in root cell elongation but are also involved in many aspects of root development, such as maintenance of meristem size, root hair formation, lateral root initiation, gravitropic response, mycorrhiza formation, and nodulation in legume species. In this review, current findings on the functions of BRs in mediating root growth, development, and symbiosis are discussed.

Negative phototropism of rice root and its influencing factors

Some characteristics of the rice (Oryza sativa L.) root were found in the experiment of unilaterally irradiating the roots which were planted in water: (i) All the seminal roots, adventitious roots and their branched roots bent away from light, and their curvatures ranged from 25° to 60°. The curvature of adventitious root of the higher node was often larger than that of the lower node, and even larger than that of the seminal root. (ii) The negative phototropic bending of the rice root was mainly due to the larger growth increment of root-tip cells of the irradiated side compared with that of the shaded side. (iii) Root cap was the site of light perception. If root cap was shaded while the root was irradiated the root showed no negative phototropism, and the root lost the characteristic of negative phototropism when root cap was divested. Rice root could resume the characteristic of negative phototropism when the new root cap grew up, if the original cells of root cap were well protected while root cap was divested, (iv) The growth increment and curvature of rice root were both influenced by light intensity. Within the range of 0-100 μmol·m-2·s-1, the increasing of light intensity resulted in the decreasing of the growth increment and the increasing of the curvature of rice root. (v) The growth increment and the curvature reached the maximum at 30℃ with the temperature treatment of 10-40℃. (vi) Blue-violet light could prominently induce the negative phototropism of rice root, while red light had no such effect. (vii) The auxin (IAA) in the solution, as a very prominent influencing factor, inhibited the growth, the negative phototropism and the gravi-tropism of rice root when the concentration of IAA increased. The response of negative phototropism of rice root disappeared when the concentration of IAA was above 10 mg·L-1.

Seven Things We Think We Know about Auxin Transport

Polar transport of the phytohormone auxin and the establishment of localized auxin maxima regulate em- bryonic development, stem cell maintenance, root and shoot architecture, and tropic growth responses. The past decade has been marked by dramatic progress in efforts to elucidate the complex mechanisms by which auxin transport regulates plant growth. As the understanding of auxin transport regulation has been increasingly elaborated, it has become clear that this process is involved in almost all plant growth and environmental responses in some way. However, we still lack information about some basic aspects of this fundamental regulatory mechanism. In this review, we present what we know （or what we think we know） and what we do not know about seven auxin-regulated processes. We discuss the role of auxin transport in gravitropism in primary and lateral roots, phototropism, shoot branching, leaf expansion, and venation. We also discuss the auxin reflux/fountain model at the root tip, flavonoid modulation of auxin transport processes, and outstanding aspects of post-translational regulation of auxin transporters. This discussion is not meant to be exhaustive, but highlights areas in which generally held assumptions require more substantive validation.