Tiller Bud Outgrowth Research Articles

Regulation of Aegilops tauschii Coss Tiller Bud Growth by Plant Density: Transcriptomic, Physiological and Phytohormonal Responses

Aegilops tauschii Coss is one of the most hazardous weeds that severely infests wheat fields in China. The tillering ability of Ae. tauschii strongly affects the occurrence and spread by influencing its seed output. In this study, Ae. tauschii was sown at low plant density (LPD) and high plant density (HPD) to investigate the effect of plant density on tiller bud outgrowth and its potential regulators using RNA-Seq. Additionally, the chlorophyll content and photosynthesis, soluble sugar and phytohormone levels were also determined at different plant densities. The results showed that an increased plant density significantly inhibited the elongation of tiller buds in the axil of the first leaf at 15 days after planting, with 7.69 mm at LPD and 1.69 mm at HPD. A total of seven putative tiller-related genes were selected and validated using quantitative real-time PCR. Furthermore, chlorophyll levels, photosynthetic efficiency, and soluble sugar contents were distinctly inhibited by HPD in Ae. tauschii, which may be responsible for the restriction of tiller bud growth. In addition, differentially expressed genes (DEGs) were markedly enriched in indole-3-acetic acid (IAA), abscisic acid (ABA), and gibberellin metabolism and signaling. Accordingly, the levels of ABA and gibberellin A3 in Ae. tauschii were strikingly higher at HPD compared with those at LPD, yet the reverse tendency was observed for IAA. Undoubtedly, such results will be highly beneficial for illuminating the underlying regulators of the Ae. tauschii tillering response to plant density and may provide new ideas for the control of this weed in the future.

Interactions Between Exogenous Cytokinin and Nitrogen Application Regulate Tiller Bud Growth via Sucrose and Nitrogen Allocation in Winter Wheat

In this study, we assessed the effects of exogenous cytokinin and nitrogen levels on sucrose and nitrogen allocation and tiller bud growth. Results showed that low nitrogen (1.25 mmol L−1, LN) inhibited tiller bud growth and decreased tiller occurrence rate, compared with high nitrogen (7.5 mmol L−1, HN). SPAD readings and leaf photosynthesis rate was decreased by LN, which resulted in the decrease of sucrose content in the tiller bud. Furthermore, LN promoted the primary and lateral root growth, which may compete with the tiller bud for sucrose and nitrogen. Application of 6-BA (30 mg L−1) increased tiller bud length and the occurrence rate. Compared with LN, leaf photosynthesis and root activity was improved by application of 6-BA under LN condition (LN + 6-BA), which caused the increase of sucrose and nitrogen accumulation in tiller bud. Moreover, LN + 6-BA significantly increased endogenous zeatin (Zt) contents and decreased abscisic acid (ABA) levels in the tiller node. Correlation analysis indicated that tiller bud growth was positively correlated with sucrose and nitrogen accumulation. The latter two were positively correlated with endogenous Zt but negatively correlated with ABA and auxin (IAA):Zt, gibberellin (GA3):Zt, and ABA:Zt. Application of 6-BA with reduced nitrogen fertilizer (240 kg N ha−1, N1 + 6-BA) increased both grain yield and the partial factor productivity of applied N. Therefore, an appropriate combination of exogenous cytokinin with reduced N application could regulate tiller growth to enhance yield by endogenous hormones balance and sucrose and nitrogen allocation in the plant.

The tin1 gene retains the function of promoting tillering in maize

Sweet maize and popcorn retain tillering growth habit during maize diversification. However, the underlying molecular genetic mechanism remains unknown. Here, we show that the retention of maize tillering is controlled by a major quantitative trait locus (QTL), tin1, which encodes a C2H2-zinc-finger transcription factor that acts independently of tb1. In sweet maize, a splice-site variant from G/GT to C/GT leads to intron retention, which enhances tin1 transcript levels and consequently increases tiller number. Comparative genomics analysis and DNA diversity analysis reveal that tin1 is under parallel selection across different cereal species. tin1 is involved in multiple pathways, directly represses two tiller-related genes, gt1 and Laba1/An-2, and interacts with three TOPLESS proteins to regulate the outgrowth of tiller buds. Our results support that maize tin1, derived from a standing variation in wild progenitor teosinte population, determines tillering retention during maize diversification.

A Trypsin Family Protein Gene Controls Tillering and Leaf Shape in Barley.

Tillering or branching is an important agronomic trait in plants, especially cereal crops. Previously, in barley (Hordeum vulgare) 'Vlamingh', we identified the high number of tillers1 (hnt1) mutant from a γ-ray-treated segregating population. hnt1 exhibited more tillers per plant, narrower leaves, and reduced plant height compared with the wild-type parent. In this study, we show that the hnt1-increased tiller number per plant is caused by accelerated outgrowth of tiller buds and that hnt1 narrower leaves are caused by a reduction in vascular tissue and cell number. Genetic analysis revealed that a 2-bp deletion in the gene HORVU2Hr1G098820 (HvHNT1), encoding a trypsin family protein, was responsible for the hnt1 mutant phenotype. Gene function was further confirmed by transgenic complementation with HvHNT1 and RNA interference experiments. HvHNT1 was expressed in vascular tissue, leaf axils, and adventitious root primordia and shown to negatively regulate tiller development. Mutation of HvHNT1 led to the accumulation of a putative cyclophilin-type peptidyl-prolyl cis/trans-isomerase (HvPPIase), which physically interacts with the HvHNT1 protein in the nucleus of plant cells. Our data suggest that HvHNT1 controls tiller development and leaf width through HvPPIase, thus contributing to understanding of the molecular players that control tillering in barley.

Gibberellic acid inhibition of tillering in tall fescue involving crosstalks with cytokinins and transcriptional regulation of genes controlling axillary bud outgrowth

Tiller production in grass species is controlled by both axillary bud initiation and bud outgrowth, which may be regulated by plant hormones. However, how gibberellic acid (GA) affects tillering in perennial grass species is still unclear. This study aims to elucidate the roles and the underlying mechanisms of GA in regulating tiller development. Tall fescue seedlings were treated with different concentrations of GA3 by foliar application, dose-dependent inhibitory effects of GA on tiller production were observed. GA3 (25 μM) slowed down the transition from axillary buds to tillers by specifically inhibiting the outgrowth of axillary buds. GA-inhibition of tillering were not related to endogenous content for auxin or strigolactone, but was mainly due to the antagonistic interaction with cytokinins (CK), as shown by the decreased CK content and up-regulation expression of CK degradation genes in GA3-treated plants. Furthermore, GA could act through regulating the expression of FaTB1 specifically expressed in axillary buds to repress bud outgrowth. These results provide insights for the regulatory mechanisms of GA for tiller bud outgrowth through crosstalks with CK and signaling of FaTB1 expression.

Tiller Bud Formation Regulators MOC1 and MOC3 Cooperatively Promote Tiller Bud Outgrowth by Activating FON1 Expression in Rice

Tillering in rice is one of the most important agronomic traits. Rice tiller development can be divided into two main processes: the formation of the axillary bud and its subsequent outgrowth. Several genes critical for bud formation in rice have been identified by genetic studies; however, their molecular functions and relationships are still largely unknown. Here, we report that MONOCULM 1 (MOC1) and MONOCULM 3/TILLERS ABSENT 1/STERILE AND REDUCED TILLERING 1 (MOC3/TAB1/SRT1), two vital regulators for tiller formation in rice, physically interact to regulate tiller bud outgrowth through upregulating the expression of FLORAL ORGAN NUMBER1 (FON1), the homolog of CLAVATA1 in rice. We found that MOC3 is able to directly bind the promoter of FON1 and subsequently activate FON1 expression. MOC1 functions as a co-activator of MOC3, whereas it could not directly bind the FON1 promoter, and further activated FON1 expression in the presence of MOC3. Accordingly, FON1 is highly expressed at axillary meristems and shows remarkably decreased expression levels in moc1 and moc3 mutants. Loss-of-function mutants of FON1 exhibit normal bud formation but defective bud outgrowth and reduced tiller number. Collectively, these results shed light on a joint transcriptional regulatory mechanim by MOC1 and MOC3, and establish a new framework for the control of tiller bud formation and outgrowth.

Sorghum tiller bud growth is repressed by contact with the overlying leaf.

Basal branching in grasses, or tillering, is an important trait determining both form and function of crops. Although similarities exist between eudicot and grass branching programs, one notable difference is that the tiller buds of grasses are covered by the subtending leaf, whereas eudicot buds are typically unconstrained. The current study shows that contact with the leaf sheath represses sorghum bud growth by providing a mechanical signal that cues the bud to refrain from rapid growth. Leaf removal resulted in massive reprogramming of the bud transcriptome that included signatures of epigenetic modifications and also implicated several hormones in the response. Bud abscisic acid transiently increased, then decreased following leaf removal relative to controls, and abscisic acid was necessary to repress bud growth in the presence of the leaf. Jasmonic acid (JA) levels and signalling increased in buds following leaf removal. Remarkably, application of JA to buds in situ promoted growth. The repression of bud growth by leaf contact shares characteristics of thigmomorphogenic responses in other systems, including the involvement of JA, though the JA effect is opposite. The repression of bud growth by leaf contact may represent a mechanism to time tillering to an appropriate developmental stage of the plant.

The Amino Acid Permease 5 (OsAAP5) Regulates Tiller Number and Grain Yield in Rice.

As fundamental nutrients, amino acids are important for rice (Oryza sativa) growth and development. Here, we identified the amino acid permease 5 (OsAAP5), that regulates tiller number and grain yield in rice. The OsAAP5 promoter sequence differed between indica and japonica rice varieties. Lower expression of OsAAP5 in the young leaf blade in indica varieties than in japonica varieties was associated with more tillers in indica than in japonica Down-regulation of OsAAP5 expression in japonica using RNA interference (RNAi) and clustered regularly interspaced short palindromic repeats led to increases in tiller number and grain yield, whereas OsAAP5 overexpression (OE) had the opposite effect. Both a protoplast amino acid uptake assay and HPLC analysis indicated that more basic (Lys, Arg) and neutral (Val, Ala) amino acids were transported and accumulated in the OE lines than in the wild type, but the opposite was observed in the RNAi lines. Furthermore, exogenous application of Lys, Arg, Val, and Ala in the OE lines substantially inhibited tiller bud elongation, but the effect was lost in the RNAi lines. Notably, concentrations of the cytokinins cis-zeatin and dihydrozeatin were much lower in the OE lines than in the wild type, whereas concentrations in the RNAi lines were higher. Thus, OsAAP5 could regulate tiller bud outgrowth by affecting cytokinin levels, and knockout of OsAAP5 could be valuable for japonica breeding programs seeking high yield and grain quality.

OsASN1 Plays a Critical Role in Asparagine-Dependent Rice Development.

Asparagine is one of the important amino acids for long-distance transport of nitrogen (N) in plants. However, little is known about the effect of asparagine on plant development, especially in crops. Here, a new T-DNA insertion mutant, asparagine synthetase 1 (asn1), was isolated and showed a different plant height, root length, and tiller number compared with wild type (WT). In asn1, the amount of asparagine decreased sharply while the total nitrogen (N) absorption was not influenced. In later stages, asn1 showed reduced tiller number, which resulted in suppressed tiller bud outgrowth. The relative expression of many genes involved in the asparagine metabolic pathways declined in accordance with the decreased amino acid concentration. The CRISPR/Cas9 mutant lines of OsASN1 showed similar phenotype with asn1. These results suggest that OsASN1 is involved in the regulation of rice development and is specific for tiller outgrowth.

Exogenous Hormonal Application Regulates the Occurrence of Wheat Tillers by Changing Endogenous Hormones.

Plant hormones play important roles in regulating the occurrence of crop tillers. However, little is known about the relationships and the underlying mechanisms between endogenous hormones and the occurrence of wheat tillers induced by exogenous hormones. In this study, two winter wheat cultivars, Xinong 979 and Xiaoyan 22, were used to investigate the effects of the exogenous application of indole-3-acetic acid (IAA) and zeatin (Z) on the occurrence of wheat tillers and investigate underlying mechanisms regulating the occurrence of tillers. The results showed that the application of IAA inhibited the occurrence of tillers, and external Z application promoted the occurrence rate of tillers under low nitrogen conditions. Further analysis of the results showed that exogenous IAA completely inhibited the growth of tiller buds, while exogenous Z significantly promoted the growth rate of tiller buds in low nitrogen conditions. Endogenous hormones exhibit important functions in regulating the growth of tiller buds, which contents were affected by exogenous hormones. Furthermore, according to the principal component analysis and correlation analysis, the growth of tiller buds was significantly positively correlated with the content of endogenous Z, whereas it was significantly negatively correlated with the ratios of endogenous IAA to endogenous Z (IAA:Z) and endogenous abscisic acid (ABA) to endogenous Z (ABA:Z). Moreover, no significant correlation was observed between the growth of the tiller buds and the endogenous IAA, endogenous gibberellins (GAs), and endogenous ABA content. These results suggested that Z played key roles in regulating the tiller occurrence, and exogenous hormones regulated the growth of wheat tiller buds via affecting the Z contents, thus regulating the occurrence of wheat tiller.

Effects of Strigolactones on Grain Yield and Seed Development in Rice

Strigolactones (SLs) are well known as a class of endogenous phytohormones that regulate tiller bud outgrowth. Reduction of inorganic phosphate (Pi) induces the accumulation of SLs, which inhibit tiller bud outgrowth in wild-type (WT) rice plants, but not in SL mutants. This suggests that SLs are important for plant adaptation to Pi-deficient conditions. Thus, we investigated the effects of SLs on grain yield and seed size in WT and dwarf (d) mutant rice plants treated with various Pi concentrations. In both WT and d mutants, plant growth, chlorophyll levels, panicle number, number of hulls, and total grain number decreased as Pi decreased, indicating that SL is not required to mediate these Pi responses. The d mutants produced more panicles than the WT control, but there was no increase in grain yield, and the seed-setting rate decreased. Removal of outgrowing tillers did not affect grain yield in d mutants. GR24 (a synthetic SL) treatment rescued grain yield in d mutants. The d3 and d53 mutants had the lowest grain yields among d mutants. Furthermore, the endosperm of d mutants was 25% smaller than that of WT plants; there were no significant differences in embryo length between WT and d mutant plants, but the endosperm cell area of the d mutants was approximately 30% smaller than that of WT plants. We propose that SLs control grain yield and rice endosperm development.

Effects of gibberellin and strigolactone on rice tiller bud growth.

Strigolactones (SLs) regulate diverse developmental phenomena. Rice SL biosynthesis and signaling mutants have an increased number of tillers and a reduced plant height relative to wild-type (WT) rice plants. In this study, we tested the effectiveness of gibberellin (GA) on restoring more tillering phenotype and dwarfism observed in both SL biosynthesis and signaling mutants. The application of GA to these mutants rescued the tiller bud outgrowth; however, the sensitivity to GA was different between the WT and the SL biosynthesis mutant.

Upregulation of DWARF27 is associated with increased strigolactone levels under sulfur deficiency in rice.

Plants produce strigolactones (SLs) in roots in response to nitrogen or phosphate deficiency. To evaluate SL levels under other mineral deficiencies in rice, we cultivated rice seedlings in hydroponic media without nitrogen, phosphorus, potassium, sulfur, calcium, magnesium, and iron. Tiller bud outgrowth was stimulated under calcium deficiency because of low SL levels. SL levels increased under sulfur deficiency, in addition to phosphate, and nitrogen deficiencies. To explore which genes are key regulators of SL production under sulfur deficiency, we analyzed the expression of SL‐related genes in sulfur‐sufficient and sulfur‐deficient conditions. An SL biosynthesis gene, DWARF27 (D27), was strongly expressed under sulfur deficiency, and its expression was decreased by sulfur supply. The levels of D10, D17, and OsMAX1 transcripts did not differ between sulfur‐sufficient and sulfur‐deficient conditions. These results suggest that the increased SL levels under sulfur deficiency are due to a high expression of D27. A combination of nitrogen, phosphorus, and sulfur deficiencies had no additive synergistic effect on SL production. Under combined phosphorus and sulfur deficiency, the expression levels of most SL biosynthesis genes were elevated. The number of tiller buds in the d27 mutant was higher than in the wild type, but lower than in other d mutants. Under sulfur deficiency, the chlorophyll content of d27 was lower than those of other d mutants. These results indicate that D27 plays an important role in adaptation to sulfur deficiency in rice.

Rice nitrate transporter OsNPF7.2 positively regulates tiller number and grain yield

BackgroundRice tiller number is one of the most important factors that determine grain yield, while nitrogen is essential for the crop growth and development, especially for tiller formation. Genes involved in nitrogen use efficiency processes have been identified in the previous studies, however, only a small number of these genes have been found to improve grain yield by promoting tillering.ResultsWe constructed over-expression (OX) lines and RNA-interference (Ri) lines, and selected a mutant of OsNPF7.2, a low-affinity nitrate transporter. Our analyses showed that rice tiller number and grain yield were significantly increased in OX lines, whereas Ri lines and mutant osnpf7.2 had fewer tiller number and lower grain yield. Under different nitrate concentrations, tiller buds grew faster in OX lines than in WT, but they grew slower in Ri lines and mutant osnpf7.2. These results indicated that altered expression of OsNPF7.2 plays a significant role in the control of tiller bud growth and regulation of tillering. Elevated expression of OsNPF7.2 also improved root length, root number, fresh weight, and dry weight. However, reduced expression of OsNPF7.2 had the opposite result on these characters. OsNPF7.2 OX lines showed more significantly enhanced influx of nitrate and had a higher nitrate concentration than WT. The levels of gene transcripts related to cytokinin pathway and cell cycle in tiller bud, and cytokinins concentration in tiller basal portion were higher in OX lines than that in WT, suggesting that altered expression of OsNPF7.2 controlled tiller bud growth and root development by regulating cytokinins content and cell cycle in plant cells. Altered expression of OsNPF7.2 also was responsible for the change in expression of the genes involved in strigolactone pathway, such as D27, D17, D10, Os900, Os1400, D14, D3, and OsFC1.ConclusionOur results suggested that OsNPF7.2 is a positive regulator of nitrate influx and concentration, and that it also regulates cell division in tiller bud and alters expression of genes involved in cytokinin and strigolactone pathways, resulting in the control over rice tiller number. Since elevated expression of OsNPF7.2 is capable of improving rice grain yield, this gene might be applied to high-yield rice breeding.

Nitrogen deficiency inhibits cell division-determined elongation, but not initiation, of rice tiller buds

Plant branching shoot number is strongly influenced by nitrogen (N) supply status. However, detailed descriptions of this phenomenon and the regulatory mechanism are lacking. In this study, we show that, in rice, in comparison to sufficient supply of ammonium sulfate or ammonium nitrate (2.5 or 5 mM N), low N (0.2 mM) or nitrate as the only N source limited shoot branching, i.e., tillering number. We observed that N deficiency did not affect the initiation, but suppressed the elongation of the tiller buds. We carried out in-situ hybridization of the tiller buds and showed that the expression of histone H4 , a marker of S-phase in the cell cycle, could not be detected in the tiller buds that had stopped growing, indicating that cell division was suppressed in the tissues. Consistent with this finding, we further detected that the expression of other cell cycle marker genes was decreased in the N-deficient tiller buds in comparison to N-sufficient tiller buds. In addition, expression of the genes involved in the strigolactone pathway was induced in the tiller buds by N deficiency, in accordance with other reports. These results shed light on the importance of proper N application to control tiller bud outgrowth in grain production.

Transcriptome Profiling of Tiller Buds Provides New Insights into PhyB Regulation of Tillering and Indeterminate Growth in Sorghum.

Phytochrome B (phyB) enables plants to modify shoot branching or tillering in response to varying light intensities and ratios of red and far-red light caused by shading and neighbor proximity. Tillering is inhibited in sorghum genotypes that lack phytochrome B (58M, phyB-1) until after floral initiation. The growth of tiller buds in the first leaf axil of wild-type (100M, PHYB) and phyB-1 sorghum genotypes is similar until 6 d after planting when buds of phyB-1 arrest growth, while wild-type buds continue growing and develop into tillers. Transcriptome analysis at this early stage of bud development identified numerous genes that were up to 50-fold differentially expressed in wild-type/phyB-1 buds. Up-regulation of terminal flower1, GA2oxidase, and TPPI could protect axillary meristems in phyB-1 from precocious floral induction and decrease bud sensitivity to sugar signals. After bud growth arrest in phyB-1, expression of dormancy-associated genes such as DRM1, GT1, AF1, and CKX1 increased and ENOD93, ACCoxidase, ARR3/6/9, CGA1, and SHY2 decreased. Continued bud outgrowth in wild-type was correlated with increased expression of genes encoding a SWEET transporter and cell wall invertases. The SWEET transporter may facilitate Suc unloading from the phloem to the apoplast where cell wall invertases generate monosaccharides for uptake and utilization to sustain bud outgrowth. Elevated expression of these genes was correlated with higher levels of cytokinin/sugar signaling in growing buds of wild-type plants.

Structure- and stereospecific transport of strigolactones from roots to shoots.

Strigolactones (SLs) are carotenoid-derived signaling molecules that mediate symbiotic and parasitic communications in the rhizosphere and plant hormones that regulate the growth and development of plants through crosstalk with other hormones. Natural SLs are classified into two groups based on the stereochemistry of the B-C ring junction. Rice and sorghum plants, both gramineous crops, produce orobanchol-type and strigol-type SLs, respectively, while tobacco plants produce both types. In the present study, we demonstrate that such species-specific phenomena in SL production also occur in the transport of exogenous SLs from roots to shoots. In rice plants, strigol-type SLs such as 5-deoxystrigol have been reported to actively inhibit tiller bud outgrowth, whereas root-applied strigol-type SLs could not be detected in shoots harvested 20 hr after treatment, indicating that metabolites of SLs or other signaling compounds downstream of SLs-but not SLs themselves-are the true inhibitors of tiller bud outgrowth.

Tillering in the sugary1 sweet corn is maintained by overriding the teosinte branched1 repressive signal

The evolution of apical dominance in maize during domestication from teosinte is associated with higher expression from the teosinte branched1 (tb1) gene that inhibits tiller bud outgrowth. Unlike many standard maize varieties, the sweet corn inbred P39 that carries a mutation in a starch biosynthesis gene sugary1 produces multiple tillers and providing an opportunity to explore the diversification of the tb1 signal in maize. Through gene expression analysis, we show that tiller buds in P39 continue to grow by overriding the high expression level of tb1 that arrests bud outgrowth in maize inbred B73. In addition, we demonstrate that while B73 is largely non-responsive to shade, both P39 and teosinte respond through tb1-independent and tb1-dependent molecular mechanisms, respectively, leading to inhibition of tiller bud outgrowth.

De novo assembly and transcriptome analysis of two contrary tillering mutants to learn the mechanisms of tillers outgrowth in switchgrass (Panicum virgatum L.).

Tillering is an important trait in monocotyledon plants. The switchgrass (Panicum virgatum), studied usually as a source of biomass for energy production, can produce hundreds of tillers in its lifetime. Studying the tillering of switchgrass also provides information for other monocot crops. High-tillering and low-tillering mutants were produced by ethyl methanesulfonate mutagenesis. Alteration of tillering ability resulted from different tiller buds outgrowth in the two mutants. We sequenced the tiller buds transcriptomes of high-tillering and low-tillering plants using next-generation sequencing technology, and generated 34 G data in total. In the de novo assembly results, 133,828 unigenes were detected with an average length of 1,238 bp, and 5,290 unigenes were differentially expressed between the two mutants, including 3,225 up-regulated genes and 2,065 down-regulated genes. Differentially expressed gene analysis with functional annotations was performed to identify candidate genes involved in tiller bud outgrowth processes using Gene Ontology classification, Cluster of Orthologous Groups of proteins, and Kyoto Encyclopedia of Genes and Genomes pathway analysis. This is the first study to explore the tillering transcriptome in two types of tillering mutants by de novo sequencing.

The interaction between nitrogen availability and auxin, cytokinin, and strigolactone in the control of shoot branching in rice (Oryza sativa L.).

Nitrogen availability and cytokinin could promote shoot branching in rice, whereas auxin and strigolactone inhibited it. The interaction between nitrogen availability and the three hormones is discussed. Rice shoot branching is strongly affected by nitrogen availability and the plant hormones auxin, cytokinin, and strigolactone; however, the interaction of them in the regulation of rice shoot branching remains a subject of debate. In the present study, nitrogen and the three hormones were used to regulate rice tiller bud growth in the indica rice variety Yangdao 6. Both nitrogen and CK promoted shoot branching in rice, whereas auxin and SL inhibited it. We used HPLC to determine the amounts of endogenous IAA and CK, and we used quantitative real-time PCR analysis to quantify the expression levels of several genes. Nitrogen enhanced the amount of CK by promoting the expression levels of OsIPTs in nodes. In addition, both nitrogen and CK downregulated the expression of genes related to SL synthesis in root and nodes, implying that the inhibition of SL synthesis by nitrogen may occur at least partially through the CK pathway. SL did not significantly reduce the amount of CK or the expression levels of OsIPT genes, but it did significantly reduce the amount of auxin and the auxin transport capacity in nodes. Auxin itself inhibited CK synthesis and promoted SL synthesis in nodes rather than in roots. Furthermore, we found that CK and SL quickly reduced and increased the expression of FC1 in buds, respectively, implying that FC1 might be a common target for the CK and SL pathways. Nitrogen and auxin delayed expression change patterns of FC1, potentially by changing the downstream signals for CK and SL.