Lamina Joint Research Articles

OsSLA1 functions in leaf angle regulation by enhancing the interaction between OsBRI1 and OsBAK1 in rice.

Leaf angle is an important trait in plants. Here, we demonstrate that the leucine-rich repeat receptor-like kinase OsSLA1 plays an important role in leaf angle regulation in rice (Oryza sativa). OsSLA1 mutant plants exhibited a small leaf angle phenotype due to changes of adaxial cells in the lamina joint. GUS staining revealed that OsSLA1 was highly expressed in adaxial cells of the lamina joint. The OsSLA1 mutant plants were insensitive to exogenous epibrassinolide (eBL) and showed upregulated expression of DWARF and CPD, but downregulated expression of BU1, BUL1, and ILI1, indicating that brassinosteroid (BR) signal transduction was blocked. Fluorescence microscopy showed that OsSLA1 was localized to the plasma membrane and nearby periplasmic vesicles. Further study showed that OsSLA1 interacts with OsBRI1 and OsBAK1 via its intracellular domain and promotes the interaction between OsBRI1 and OsBAK1. In addition, phosphorylation experiments revealed that OsSLA1 does not possess kinase activity, but that it can be phosphorylated by OsBRI1 in vitro. Knockout of OsSLA1 in the context of d61 caused exacerbation of the mutant phenotype. These results demonstrate that OsSLA1 regulates leaf angle formation via positive regulation of BR signaling by enhancing the interaction of OsBRI1 with OsBAK1.

Rice OsIAA6 interacts with OsARF1 and regulates leaf inclination

Leaf inclination, a component of crop architecture, influences photosynthetic efficiency and planting density. Various factors, particularly the phytohormones auxin and brassinosteroids (BRs), function in regulating lamina joint bending, and understanding of the genetic control of leaf inclination will help to elucidate the relevant regulatory network. Screening a rice T-DNA insertion population revealed a mutant that was insensitive to auxin and displayed an enlarged leaf angle due to increased cell length on the adaxial side of the lamina joint. Genetic analysis revealed that the increased leaf inclination was caused by T-DNA insertion in the promoter region of OsIAA6, resulting in elevated OsIAA6 expression. Further study showed that OsIAA6 interacts with OsARF1 to suppress auxin signaling and regulates leaf inclination. OsIAA6 mediates the BR effects on lamina joint development, and OsBZR1, the key transcription factor in BR signaling, binds directly to the promoter of OsIAA6 to stimulate its transcription. These results indicate the roles of the OsIAA6–OsARF1 module in regulating rice leaf inclination and suggest the synergistic effects of the phytohormones auxin and BR.

Transcription factor OsbZIP49 controls tiller angle and plant architecture through the induction of indole-3-acetic acid-amido synthetases in rice.

Tiller angle is an important determinant of plant architecture in rice (Oryza sativa L.). Auxins play a critical role in determining plant architecture; however, the underlying metabolic and signaling mechanisms are still largely unknown. In this study, we have identified a member of the bZIP family of TGA class transcription factors, OsbZIP49, that participates in the regulation of plant architecture and is specifically expressed in gravity-sensing tissues, including the shoot base, nodes and lamina joints. Transgenic rice plants overexpressing OsbZIP49 displayed a tiller-spreading phenotype with reduced plant height and internode lengths. In contrast, CRISPR/Cas9-mediated knockout of OsbZIP49 resulted in a compact architecture. Follow-up studies indicated that the effects of OsbZIP49 on tiller angles are mediated through changes in shoot gravitropic responses. Additionally, we provide evidence that OsbZIP49 activates the expression of indole-3-acetic acid-amido synthetases OsGH3-2 and OsGH3-13 by directly binding to TGACG motifs located within the promoters of both genes. Increased GH3-catalyzed conjugation of indole-3-acetic acid (IAA) in rice transformants overexpressing OsbZIP49 resulted in the increased accumulation of IAA-Asp and IAA-Glu, and a reduction in local free auxin, tryptamine and IAA-Glc levels. Exogenous IAA or naphthylacetic acid (NAA) partially restored shoot gravitropic responses in OsbZIP49-overexpressing plants. Knockout of OsbZIP49 led to reduced expression of both OsGH3-2 and OsGH3-13 within the shoot base, and increased accumulation of IAA and increased OsIAA20 expression levels were observed in transformants following gravistimulation. Taken together, the present results reveal the role transcription factor OsbZIP49 plays in determining plant architecture, primarily due to its influence on local auxin homeostasis.

Modulation of Rice Leaf Angle and Grain Size by Expressing OsBCL1 and OsBCL2 under the Control of OsBUL1 Promoter.

Leaf angle and grain size are important agronomic traits affecting rice productivity directly and/or indirectly through modulating crop architecture. OsBC1, as a typical bHLH transcription factor, is one of the components comprising a complex formed with LO9-177 and OsBUL1 contributing to modulation of rice leaf inclination and grain size. In the current study, two homologues of OsBC1, OsBCL1 and OsBCL2 were functionally characterized by expressing them under the control of OsBUL1 promoter, which is preferentially expressed in the lamina joint and the spikelet of rice. Increased leaf angle and grain length with elongated cells in the lamina joint and the grain hull were observed in transgenic rice containing much greater gibberellin A3 (GA3) levels than WT, demonstrating that both OsBCL1 and OsBCL2 are positive regulators of cell elongation at least partially through increased GA biosynthesis. Moreover, the cell elongation was likely due to cell expansion rather than cell division based on the related gene expression and, the cell elongation-promoting activities of OsBCL1 and OsBCL2 were functional in a dicot species, Arabidopsis.

AUXIN RESPONSE FACTORS 6 and 17 control the flag leaf angle in rice by regulating secondary cell wall biosynthesis of lamina joints.

Flag leaf angle impacts the photosynthetic capacity of densely grown plants and is thus an important agronomic breeding trait for crop architecture and yield. The hormone auxin plays a key role in regulating this trait, yet the underlying molecular and cellular mechanisms remain unclear. Here, we report that two rice (Oryza sativa) auxin response factors (ARFs), OsARF6 and OsARF17, which are highly expressed in lamina joint tissues, control flag leaf angle in response to auxin. Loss-of-function double osarf6 osarf17 mutants displayed reduced secondary cell wall levels of lamina joint sclerenchymatous cells (Scs), resulting in an exaggerated flag leaf angle and decreased grain yield under dense planting conditions. Mechanical measurements indicated that the mutant lamina joint tissues were too weak to support the weight of the flag leaf blade, resembling the phenotype of the rice increased leaf angle1 (ila1) mutant. We demonstrate that OsARF6 and OsARF17 directly bind to the ILA1 promoter independently and synergistically to activate its expression. In addition, auxin-induced ILA1 expression was dependent on OsARF6 and OsARF17. Collectively, our study reveals a mechanism that integrates auxin signaling with the secondary cell wall composition to determine flag leaf angle, providing breeding targets in rice, and potentially other cereals, for this key trait.

Leaf direction: Lamina joint development and environmental responses.

Plant architecture plays a major role in canopy photosynthesis and biomass production, and plants adjust their growth (and thus architecture) in response to changing environments. Leaf angle is one of the most important traits in rice (Oryza sativa L.) plant architecture, because leaf angle strongly affects leaf direction and rice production, with more-erect leaves being advantageous for high-density plantings. The degree of leaf bending depends on the morphology of the lamina joint, which connects the leaf and the sheath. In this review, we discuss cell morphology in different lamina joint tissues and describe the underlying genetic network that governs this morphology and thus regulates leaf direction. Furthermore, we focus on the mechanism by how environmental factors influence rice leaf angle. Our review provides a theoretical framework for the future genetic improvement of rice leaf orientation and plant architecture.

Quantitative trait loci for leaf inclination angle in rice detected using reciprocal mapping populations

• Six Takanari alleles increased leaf inclination angle at least twice in four years. • No Koshihikari alleles consistently increased leaf inclination angle. • Takanari alleles decreased the length of lamina joint cells on the adaxial side. • Takanari alleles improved light penetration into the canopy. • The effects of Takanari alleles were not enhanced by pyramiding QTLs. The improvement of canopy architecture is a feasible way to increase rice productivity. However, its genetic control in rice cultivars in the field is not well understood. The high-yielding indica cultivar Takanari has more erect leaves (a larger leaf inclination angle: LIA) during ripening, and this contributes to its higher dry matter production than that of the widely grown but less productive japonica cultivar Koshihikari. To identify quantitative trait loci (QTLs) for increasing LIA, we used reciprocal mapping populations of backcross inbred lines (131 lines in 2009 and 143 lines in 2010) and chromosome segment substitution lines (80 lines in 2011 and 2012) derived from a cross between Takanari and Koshihikari. Six Takanari alleles increased LIA at least in two of the four years, but we could not detect any such Koshihikari alleles. Five QTLs for LIA were expressed in plants with both Koshihikari and Takanari genetic backgrounds. LIA of two near isogenic lines carrying only one Takanari allele reached the level of Takanari. The Takanari alleles of the QTLs decreased the length of parenchyma cells in the lamina joint on the adaxial side in plants with the Koshihikari background, decreasing the ratio of the lamina joint length on the adaxial side to that on the abaxial side (LJ ratio) and thus increasing LIA. Three Takanari alleles increased dry matter production in plants with Koshihikari background via decreasing the canopy extinction coefficient ( k ). However, any pyramiding effects of Takanari alleles on LJ ratio, LIA, k and dry matter production were not observed.

OsbHLH98 regulates leaf angle in rice through transcriptional repression of OsBUL1

Leaf angle is an important agronomic trait in cereals that helps determine plant yield by affecting planting density. However, the regulation mechanism of leaf angle remained elusive. Here, we show that OsbHLH98, a rice bHLH transcription factor, negatively regulates leaf angle. osbhlh98 mutant leaves formed a larger leaf angle, whereas transgenic plants overexpressing OsbHLH98 exhibited a slight reduction in leaf angle. We determined that the changes in leaf angle resulted from increased number and size of parenchyma cells on the adaxial side of the lamina joint in osbhlh98 mutants. Experiments using reporter constructs showed that OsbHLH98 is expressed on the adaxial side of lamina joints, consistent with its proposed function in regulating leaf angle. Furthermore, we established by chromatin immunoprecipitation and CUT&RUN that OsBUL1 is a direct downstream target of OsbHLH98. Transactivation assays and reverse transcription quantitative polymerase chain reaction (RT-qPCR) analysis indicated that OsbHLH98 represses OsBUL1 transcription. Our results demonstrate that OsbHLH98 negatively regulates leaf angle by counteracting brassinosteroid-induced cell elongation via the repression of OsBUL1 transcription. The characterization of OsbHLH98 and its role in determining leaf angle will lay the foundation to develop the ideal plant architecture for adaptation to high planting density.

Effects of 24-Epibrassinolide, Bikinin, and Brassinazole on Barley Growth under Salinity Stress Are Genotype- and Dose-Dependent

Brassinosteroids (BRs) are involved in the regulation of many plant developmental processes and stress responses. In the presented study, we found a link between plant growth under salinity stress and sensitivity to 24-epibrassinolide (24-EBL, the most active phytohormone belonging to BRs), brassinazole (Brz) and bikinin (inhibitors of BR biosynthesis and signaling pathways, respectively). Plant sensitivity to treatment with active substances and salinity stress was genotype-dependent. Cv. Haruna Nijo was more responsive during the lamina joint inclination test, and improved shoot and root growth at lower concentrations of 24-EBL and bikinin under salinity stress, while cv. Golden Promise responded only to treatments of higher concentration. The use of Brz resulted in significant dose-dependent growth inhibition, greater for cv. Haruna Nijo. The results indicated that BR biosynthesis and/or signaling pathways take part in acclimation mechanisms, however, the regulation is complex and depends on internal (genotypic and tissue/organ sensitivity) and external factors (stress). Our results also confirmed that the lamina joint inclination test is a useful tool to define plant sensitivity to BRs, and to BR-dependent salinity stress. The test can be applied to manipulate the growth and stress responses of crops in agricultural practice or to select plants that are sensitive/tolerant to salinity stress in the plant breeding projects.

Spatiotemporal Resolved Leaf Angle Establishment Improves Rice Grain Yield via Controlling Population Density.

SummaryLeaf angle is mainly determined by the lamina joint (LJ) and contributes to ideal crop architecture for high yield. Here, we dissected five successive stages with distinct cytological features of LJs spanning organogenesis to leaf angle formation and obtained the underlying stage-specific mRNAs and small RNAs, which well explained the cytological dynamics during LJ organogenesis and leaf angle plasticity. Combining the gene coexpression correlation with high-throughput promoter analysis, we identified a set of transcription factors (TFs) determining the stage- and/or cytological structure-specific profiles. The functional studies of these TFs demonstrated that cytological dynamics determined leaf angle and that the knockout rice of these TFs with erect leaves significantly enhanced yield by maintaining the proper tiller number under dense planting. This work revealed the high-resolution mechanisms of how the cytological dynamics of LJ determined leaf erectness and served as a valuable resource to remodel rice architecture for high yield by controlling population density.

Physiological and transcriptomic analyses of brassinosteroid function in moso bamboo (Phyllostachys edulis) seedlings.

This study demonstrates that brassinosteroid is essential for seedling and shoot growth in moso bamboo. The shoot of moso bamboo is known to grow extremely fast. The roles of phytohormones in such fast growth of bamboo shoot remain unclear. Here we reported that endogenous brassinosteroid (BR) is a major factor promoting bamboo shoot internode elongation. Reducing endogenous brassinosteroid level by its biosynthesis inhibitor propiconazole stunted shoot growth in seedling stage, whereas exogenous BR application promoted scale leaf elongation and the inclination of lamina joint of leaves and scale leaves. Genome-wide transcriptome analysis identified hundreds of genes whose expression levels are altered by BR and propiconazole in shoots and roots of bamboo seedling. The data show that BR regulates cell wall-related genes, hydrogen peroxide catabolic genes, and auxin-related genes. Our study demonstrates an essential role of BR in fast growth bamboo shoots and identifies a large number of BR-responsive genes in bamboo seedlings.

Rice GROWTH-REGULATING FACTOR7 Modulates Plant Architecture through Regulating GA and Indole-3-Acetic Acid Metabolism.

Plant-specific GROWTH-REGULATING FACTORs (GRFs) participate in central developmental processes, including leaf and root development; inflorescence, flower, and seed formation; senescence; and tolerance to stresses. In rice (Oryza sativa), there are 12 GRFs, but the role of the miR396-OsGRF7 regulatory module remains unknown. Here, we report that OsGRF7 shapes plant architecture via the regulation of auxin and GA metabolism in rice. OsGRF7 is mainly expressed in lamina joints, nodes, internodes, axillary buds, and young inflorescences. Overexpression of OsGRF7 causes a semidwarf and compact plant architecture with an increased culm wall thickness and narrowed leaf angles mediated by shortened cell length, altered cell arrangement, and increased parenchymal cell layers in the culm and adaxial side of the lamina joints. Knockout and knockdown lines of OsGRF7 exhibit contrasting phenotypes with severe degradation of parenchymal cells in the culm and lamina joints at maturity. Further analysis indicated that OsGRF7 binds the ACRGDA motif in the promoters of a cytochrome P450 gene and AUXIN RESPONSE FACTOR12, which are involved in the GA synthesis and auxin signaling pathways, respectively. Correspondingly, OsGRF7 alters the contents of endogenous GAs and auxins and sensitivity to exogenous phytohormones. These findings establish OsGRF7 as a crucial component in the OsmiR396-OsGRF-plant hormone regulatory network that controls rice plant architecture.

The \u201cslide technique\u201d\u2014a novel free-hand method of subaxial cervical pedicle screw placement

BackgroundCervical Pedicle Screw (CPS) placement is a challenging work due to the high risk of neurovascular complications. Although there have been several different free-hand or navigation assisted techniques for CPS placement, perforations may occur during screw insertion, especially lateral perforation. The objective of this manuscript is to describe a novel free-hand technique for subaxial CPS placement (C3–C7) and to evaluate if it decreases the chances of perforation.MethodsThirty-two patients undergoing surgery with CPS instrumentation (C3–C7) at our institute between June 2017 and December 2018 were included in this study. All the patients had cervical trauma, and pedicle screw insertion was performed according to the free-hand “slide technique”. The lamina, lateral mass and facet joint of the target area were exposed and the optimal entry point was found on the lateral mass posterior surface. A pedicular probe was then inserted and gently advanced. During the pedicle probe insertion, the cortex of the medial margin of the pedicle acted as a slide to permit the safe insertion of the screw. If the pedicle screw pathway was intact, the screw of the appropriate size was carefully placed. Three-dimensional (3D) CT imaging reconstruction was performed in all the patients after surgery, and screw perforations were graded with the Gertzbein-Robbins classification.ResultsThirty-two patients who met the inclusion criteria were included in this study. A total of 257 CPSs (C3–7) were inserted, of which 41 CPSs were in C3, 61 CPSs were in C4, 55 CPSs were in C5, 53 CPSs were in C6, and 47 CPSs were in C7. The diameter and length of CPSs were 3.5 mm and 22–26 mm respectively. According to the Gertzbein-Robbins classification, grade 0, 231 screws; grade 1, 19 screws; and grade 2, 7 screws. No neurovascular complications occurred stemming from malpositioning of pedicle screws. Among perforated screws (26 screws), there were 16 lateral perforations, 5 medical perforations, and 4 inferior perforations.ConclusionsThe initial usage result shows the “slide technique” is a safe, effective and cost-effective technique for pedicle screw placement in the cervical spine. This is the first report of such a technique, and further studies are needed.

The molecular basis of the interaction between Brassinosteroid induced and phosphorous deficiency induced leaf inclination in rice

The phosphorous deficiency in arable land limits crop production globally. Plants developed a set of coordinated biochemical and developmental responses to cope with Pi deficiency during evolution. One of typical developmental responses to Pi deficiency is the induction of leaf erectness, which reduced the light capture ability and inhibited photosynthesis to conserve Pi in rice. It has been revealed that Pi deficiency induced leaf inclination by regulating the expression of BR pathway genes. However, how canonic BR signaling coordinates Pi deficiency responses in rice lamina joint development was not clear. Understanding mechanism underlying Pi-deficiency-induced leaf inclination enable us to breed new rice cultivars with increased Pi efficiency. Here we reported the molecular mechanism underlying the interaction of phosphorous deficiency-induced and BR-induced leaf inclination. We showed that BR deficiency can attenuate the leaf inclination by compromising Pi deficiency-induced BU1 expression and that constitutively activated or repressed BR signaling resulted in the insensitivity of Pi deficiency-induced leaf inclination. Furthermore, we compared expression profile of WT and BR signaling constitutively activated or repressed transgenic plants under normal and deficient phosphorous conditions by RNA-seq analysis. Our work revealed the complexity of Pi deficiency stress-induced and BR induced leaf inclinations in rice.

The Rice Basic Helix-Loop-Helix 79 (OsbHLH079) Determines Leaf Angle and Grain Shape.

Changes in plant architecture, such as leaf size, leaf shape, leaf angle, plant height, and floral organs, have been major factors in improving the yield of cereal crops. Moreover, changes in grain size and weight can also increase yield. Therefore, screens for additional factors affecting plant architecture and grain morphology may enable additional improvements in yield. Among the basic Helix-Loop-Helix (bHLH) transcription factors in rice (Oryza sativa), we found an enhancer-trap T-DNA insertion mutant of OsbHLH079 (termed osbhlh079-D). The osbhlh079-D mutant showed a wide leaf angle phenotype and produced long grains, similar to the phenotypes of mutants with increased brassinosteroid (BR) levels or enhanced BR signaling. Reverse transcription-quantitative PCR analysis showed that BR signaling-associated genes are largely upregulated in osbhlh079-D, but BR biosynthesis-associated genes are not upregulated, compared with its parental japonica cultivar ‘Dongjin’. Consistent with this, osbhlh079-D was hypersensitive to BR treatment. Scanning electron microscopy revealed that the expansion of cell size in the adaxial side of the lamina joint was responsible for the increase in leaf angle in osbhlh079-D. The expression of cell-elongation-associated genes encoding expansins and xyloglucan endotransglycosylases/hydrolases increased in the lamina joints of leaves in osbhlh079-D. The regulatory function of OsbHLH079 was further confirmed by analyzing 35S::OsbHLH079 overexpression and 35S::RNAi-OsbHLH079 gene silencing lines. The 35S::OsbHLH079 plants showed similar phenotypes to osbhlh079-D, and the 35S::RNAi-OsbHLH079 plants displayed opposite phenotypes to osbhlh079-D. Taking these observations together, we propose that OsbHLH079 functions as a positive regulator of BR signaling in rice.

Strigolactones Decrease Leaf Angle in Response to Nutrient Deficiencies in Rice.

Strigolactones (SLs) are a class of plant hormones that are synthesized from β-carotene through sequential reactions catalyzed by DWARF (D) 27, D17, D10, and OsMORE AXILLARY GROWTH (MAX) 1 in rice (Oryza sativa L.). In rice, endogenous SL levels increase in response to deficiency of nitrogen, phosphate, or sulfate (−N, −P, or −S). Rice SL mutants show increased lamina joint (LJ) angle as well as dwarfism, delayed leaf senescence, and enhanced shoot branching. The LJ angle is an important trait that determines plant architecture. To evaluate the effect of endogenous SLs on LJ angle in rice, we measured LJ angle and analyzed the expression of SL-biosynthesis genes under macronutrient deficiencies. In the “Shiokari” background, LJ angle was significantly larger in SL mutants than in the wild-type (WT). In WT and SL-biosynthesis mutants, direct treatment with the SL synthetic analog GR24 decreased the LJ angle. In WT, deficiency of N, P, or S, but not of K, Ca, Mg, or Fe decreased LJ angle. In SL mutants, deficiency of N, P, or S had no such effect. We analyzed the time course of SL-related gene expression in the LJ of WT deficient in N, P, or S, and found that expression of SL-biosynthesis genes increased 2 or 3 days after the onset of deficiency. Expression levels of both the SL-biosynthesis and signaling genes was particularly strongly increased under −P. Rice cultivars “Nipponbare”, “Norin 8”, and “Kasalath” had larger LJ angle than “Shiokari”, interestingly with no significant differences between WT and SL mutants. In “Nipponbare”, endogenous SL levels increased and the LJ angle was decreased under −N and −P. These results indicate that SL levels increased in response to nutrient deficiencies, and that elevated endogenous SLs might negatively regulate leaf angle in rice.

The basic helix-loop-helix transcription factor OsBLR1 regulates leaf angle in rice via brassinosteroid signalling.

Leaf angle is a key factor in plant architecture and crop yield. Brassinosteroids (BRs) regulate many developmental processes, especially the leaf angle in monocots. However, the BR signalling pathway is complex and includes many unknown members. Here, we propose that Oryza sativa BRASSINOSTEROID-RESPONSIVE LEAF ANGLE REGULATOR 1 (OsBLR1) encodes a bHLH transcription factor, and positively regulates BR signalling to increase the leaf angle and grain length in rice (Oryza sativa L.). Lines overexpressing OsBLR1 (blr1-D and BLR1-OE-1/2/3) had similar traits, with increased leaf angle and grain length. Conversely, OsBLR1-knockout mutants (blr1-1/2/3) had erect leaves and shorter grains. Lamina joint inclination, coleoptile elongation, and root elongation assay results indicated that these overexpression lines were more sensitive to BR, while the knockout mutants were less sensitive. There was no significant difference in the endogenous BR contents of blr1-1/2 and wild-type plants. These results suggest that OsBLR1 is involved in BR signal transduction. The blr1-D mutant, with increased cell growth in the lamina joint and smaller leaf midrib, showed significant changes in gene expression related to the cell wall and leaf development compared with wild-type plants; furthermore, the cellulose and protopectin contents in blr1-D were reduced, which resulted in the increased leaf angle and bent leaves. As the potential downstream target gene of OsBLR1, the REGULATOR OF LEAF INCLINATION1 (OsRLI1) gene expression was up-regulated in OsBLR1-overexpression lines and down-regulated in OsBLR1-knockout mutants. Moreover, we screened OsRACK1A as an interaction protein of OsBLR1 using a yeast two-hybrid assay and glutathione-S-transferase pull-down.

Semi-dwarf 1 (sd1) gene enhances light penetration into the canopy through regulating leaf inclination angle in rice

Manipulating leaf inclination angle is a feasible approach to improve canopy photosynthesis and dry matter production in rice (Oryza sativa) via enhancing light penetration into the canopy. The importance of the semi-dwarf 1 (sd1) gene, a loss-of-function allele of SD1 encoding GA-20 oxidase-2 in gibberellin (GA) biosynthesis, in rice yield through improving lodging resistance is well known. However, little is known about the effects of sd1 as well as GA on leaf inclination angle. A near-isogenic line carrying the ‘Takanari’ sd1 allele in the ‘Koshihikari’ genetic background (NIL-sd1) had leaves with larger (more erect) leaf inclination angle and showed a smaller canopy extinction coefficient at the ripening stage than did ‘Koshihikari’. Furthermore, the leaf inclination angle of ‘Reimei’, an sd1 mutant in the ‘Fujiminori’ background, and of several other sd1 mutants in ‘Koshihikari’ background, was larger at the ripening stage than in parental lines, ‘Fujiminori’ and ‘Koshihikari’, respectively. The flag leaves of ‘Koshihikari’ plants treated with a GA biosynthesis inhibitor, paclobutrazol (PBZ), displayed a larger leaf inclination angle than did those of mock-treated plants during ripening. Treatment at the lamina joint of a flag leaf with exogenous GA3 or brassinolide (BL) significantly decreased leaf inclination angle. BL treatment increased the thickness of lamina joint tissue and the longitudinal cell length on the adaxial side of the marginal region of lamina joint, whereas GA3 treatment only increased the longitudinal cell length. NIL-sd1 had a lamina joint with decreased longitudinal adaxial cell length, but no difference in abaxial cell length nor tissue thickness in lamina joint cross section compared with that of ‘Koshihikari’. Thus, sd1 regulates the leaf inclination angle via a GA-based mechanism but not via BL involved in the crosstalk with GA.

Abscisic Acid Represses Rice Lamina Joint Inclination by Antagonizing Brassinosteroid Biosynthesis and Signaling.

Leaf angle is a key parameter that determines plant architecture and crop yield. Hormonal crosstalk involving brassinosteroid (BR) plays an essential role in leaf angle regulation in cereals. In this study, we investigated whether abscisic acid (ABA), an important stress-responsive hormone, co-regulates lamina joint inclination together with BR, and, if so, what the underlying mechanism is. Therefore, lamina joint inclination assay and RNA sequencing (RNA-Seq) analysis were performed here. ABA antagonizes the promotive effect of BR on leaf angle. Hundreds of genes responsive to both hormones that are involved in leaf-angle determination were identified by RNA-Seq and the expression of a gene subset was confirmed using quantitative real-time PCR (qRT-PCR). Results from analysis of rice mutants or transgenic lines affected in BR biosynthesis and signaling indicated that ABA antagonizes the effect of BR on lamina joint inclination by targeting the BR biosynthesis gene D11 and BR signaling genes GSK2 and DLT, thus forming a multi-level regulatory module that controls leaf angle in rice. Taken together, our findings demonstrate that BR and ABA antagonistically regulate lamina joint inclination in rice, thus contributing to the elucidation of the complex hormonal interaction network that optimizes leaf angle in rice.

Wheat TaSPL8 Modulates Leaf Angle Through Auxin and Brassinosteroid Signaling.

In grass crops, leaf angle is determined by development of the lamina joint, the tissue connecting the leaf blade and sheath, and is closely related to crop architecture and yield. In this study, we identified a mutant generated by fast neutron radiation that exhibited an erect leaf phenotype caused by defects in lamina joint development. Map-based cloning revealed that the gene TaSPL8, encoding a SQUAMOSA PROMOTER BINDING-LIKE (SPL) protein, is deleted in this mutant. TaSPL8 knock-out mutants exhibit erect leaves due to loss of the lamina joint, compact architecture, and increased spike number especially in high planting density, suggesting similarity with its LIGULESS1 homologs in maize (Zea mays) and rice (Oryza sativa). Hence, LG1 could be a robust target for plant architecture improvement in grass species. Common wheat (Triticum aestivum, 2n = 6× = 42; BBAADD) is an allohexaploid containing A, B, and D subgenomes and the homeologous gene of TaSPL8 from the D subgenome contributes to the length of the lamina joint to a greater extent than that from the A and B subgenomes. Comparison of the transcriptome between the Taspl8 mutant and the wild type revealed that TaSPL8 is involved in the activation of genes related to auxin and brassinosteroid pathways and cell elongation. TaSPL8 binds to the promoters of the AUXIN RESPONSE FACTOR gene and of the brassinosteroid biogenesis gene CYP90D2 and activates their expression. These results indicate that TaSPL8 might regulate lamina joint development through auxin signaling and the brassinosteroid biosynthesis pathway.