Regulating Plant Architecture Research Articles

Identification of a candidate dwarfing gene in Pallas, the first commercial barley cultivar generated through mutational breeding.

Many induced mutants are available in barley (Hordeum vulgare L.). One of the largest groups of induced mutants is the Erectoides (ert) mutants, which is characterized by a compact and upright spike and a shortened culm. One isolated mutant, ert-k.32, generated by X-ray treatment and registered in 1958 under the named "Pallas", was the first ever induced barley mutant to be released on the market. Its value was improved culm strength and enhanced lodging resistance. In this study, we aimed to identify the casual gene of the ert-k.32 mutant by whole genome sequencing of allelic ert-k mutants. The suggested Ert-k candidate gene, HORVU.MOREX.r3.6HG0574880, is located in the centromeric region of chromosome 6H. The gene product is an alpha/beta hydrolase with a catalytic triad in the active site composed of Ser-167, His-261 and Asp-232. In comparison to proteins derived from the Arabidopsis genome, ErtK is most similar to a thioesterase with de-S-acylation activity. This suggests that ErtK catalyzes post-translational modifications by removing fatty acids that are covalently attached to cysteine residues of target proteins involved in regulation of plant architecture and important commercial traits such as culm stability and lodging resistance.

Dysfunction of an Anaphase-Promoting Complex Subunit 8 Homolog Leads to Super-Short Petioles and Enlarged Petiole Angles in Soybean

Plant height, petiole length, and the angle of the leaf petiole and branch angles are crucial traits determining plant architecture and yield in soybean (Glycine max L.). Here, we characterized a soybean mutant with super-short petioles (SSP) and enlarged petiole angles (named Gmssp) through phenotypic observation, anatomical structure analysis, and bulk sequencing analysis. To identify the gene responsible for the Gmssp mutant phenotype, we established a pipeline involving bulk sequencing, variant calling, functional annotation by SnpEFF (v4.0e) software, and Integrative Genomics Viewer analysis, and we initially identified Glyma.11G026400, encoding a homolog of Anaphase-promoting complex subunit 8 (APC8). Another mutant, t7, with a large deletion of many genes including Glyma.11G026400, has super-short petioles and an enlarged petiole angle, similar to the Gmssp phenotype. Characterization of the t7 mutant together with quantitative trait locus mapping and allelic variation analysis confirmed Glyma.11G026400 as the gene involved in the Gmssp phenotype. In Gmssp, a 4 bp deletion in Glyma.11G026400 leads to a 380 aa truncated protein due to a premature stop codon. The dysfunction or absence of Glyma.11G026400 caused severe defects in morphology, anatomical structure, and physiological traits. Transcriptome analysis and weighted gene co-expression network analysis revealed multiple pathways likely involved in these phenotypes, including ubiquitin-mediated proteolysis and gibberellin-mediated pathways. Our results demonstrate that dysfunction of Glyma.11G026400 leads to diverse functional consequences in different tissues, indicating that this APC8 homolog plays key roles in cell differentiation and elongation in a tissue-specific manner. Deciphering the molecular control of petiole length and angle enriches our knowledge of the molecular network regulating plant architecture in soybean and should facilitate the breeding of high-yielding soybean cultivars with compact plant architecture.

Conformational Dynamics of the D53-D3-D14 Complex in Strigolactone Signaling.

Strigolactones (SLs) play fundamental roles in regulating plant architecture, which is a major factor determining crop yield. The perception and signal transduction of SLs require the formation of a complex containing the receptor DWARF14 (D14), an F-box protein D3 and a transcriptional regulator D53 in an SL-dependent manner. Structural and biochemical analyses of D14 and its orthologues DAD2 and AtD14, D3, and the complexes of ASK1-D3-AtD14 and D3CTH-D14 have made great contributions to understanding the mechanisms of SL perception. However, structural analyses of D53 and the D53-D3-D14 holo-complex are challenging, and the biochemical mechanism underlying complex assembly remains poorly understood. Here, we found that apo-D53 was rather flexible and reconstituted the holo-complex containing D53, SKP1, D3, and D14 with rac-GR24. The cryo-EM structure of SKP1-D3-D14 in the presence of D53 was analyzed and superimposed on the crystal structure of ASK1-D3-AtD14 without D53. No large conformational rearrangement was observed but a 9 Å rotation appeared between D14 and AtD14. Using hydrogen-deuterium exchange monitored by mass spectrometry, we analyzed dynamic motifs of D14, D3 and D53 in the D53-SKP1-D3-D14 complex assembly process and further identified two potential interfaces in D53 that are located in the N and D2 domains, respectively. Together, our results uncovered the dynamic conformational changes and built a model of the holo-complex D53-SKP1-D3-D14, offering valuable information for the biochemical and genetic mechanisms of SL perception and signal transduction.

Different Concentrations of Chemical Topping Agents Affect Cotton Yield and Quality by Regulating Plant Architecture

Manual removal of the main stem tips of cotton (Gossypium hirsutum L.) is a traditional topping approach in China. However, chemical topping has become an inevitable trend. Therefore, it is of great significance to study the effect and appropriate concentration of agents for advancement of cotton whole process mechanization management technology. A two-year field experiment was conducted to evaluate the effects of different methods of topping on cotton yield and plant architecture in Shihezi, Xinjiang, China. Plant topping included manual topping, non-topping as the control, and chemical topping (high, medium, and low concentrations of topping agent) to determine a suitable topping method and topping agent concentration for machine-picked cotton. Chemical topping was performed using fortified mepiquat chloride (DPC+). Chemical topping and manual topping increased seed cotton yield compared with non-topping. Compared with non-topping, high, medium, and low concentrations of topping agent significantly increased the yield by 19.5–27.9%, 24.1–29.4%, and 24.3–28.4%, respectively. Topping treatment mainly regulated yield and total boll number per unit area by affecting the boll number per plant and had a certain positive effect on fiber strength but no significant effect on boll weight. Chemical topping affected both vertical and horizontal plant architecture characteristics of cotton; the plant height of low-, medium-, and high-concentration treatments increased by 7.2–11.4 cm, 4.0–5.7 cm, and 2.3–4.4 cm, respectively, compared with manual topping and decreased by 5.1–7.8 cm, 8.3–13.5 cm, and 9.4–16.9 cm, respectively, compared with non-topping. The number of main stem internodes was significantly different between high- and low-concentration treatments, which showed that the higher the concentration, the lower the number of the main stem internodes. Chemical topping controlled the increase in the length of the upper branches, the higher the concentration, the lower the increase in the length of the upper fruit branch. Compared with non-topping, the medium concentration of topping agent controlled the number of fruit branches, similar to manual topping. The role of upper internode length of cotton cannot be ignored under chemical topping. The peak leaf area index (LAI) of medium- and high-concentration treatments was delayed for 1 week in the late growth period (after topping for 28 d) compared with manual topping. The LAI values were high, and the duration of high values was prolonged. The optimal chemical topping agent was the medium concentration.

Photoperiod and gravistimulation-associated Tiller Angle Control 1 modulates dynamic changes in rice plant architecture.

TAC1 is involved in photoperiodic and gravitropic responses to modulate rice dynamic plant architecture likely by affecting endogenous auxin distribution, which could explain TAC1 widespread distribution in indica rice. Plants experience a changing environment throughout their growth, which requires dynamic adjustments of plant architecture in response to these environmental cues. Our previous study demonstrated that Tiller Angle Control 1 (TAC1) modulates dynamic changes in plant architecture in rice; however, the underlying regulatory mechanisms remain largely unknown. In this study, we show that TAC1 regulates plant architecture in an expression dose-dependent manner, is highly expressed in stems, and exhibits dynamic expression in tiller bases during the growth period. Photoperiodic treatments revealed that TAC1 expression shows circadian rhythm and is more abundant during the dark period than during the light period and under short-day conditions than under long-day conditions. Therefore, it contributes to dynamic plant architecture under long-day conditions and loose plant architecture under short-day conditions. Gravity treatments showed that TAC1 is induced by gravistimulation and negatively regulates shoot gravitropism, likely by affecting auxin distribution. Notably, the tested indica rice containing TAC1 displayed dynamic plant architecture under natural long-day conditions, likely explaining the widespread distribution of TAC1 in indica rice. Our results provide new insights into TAC1-mediated regulatory mechanisms for dynamic changes in rice plant architecture.

Genetic mapping identified three hotspot genomic regions and candidate genes controlling heat tolerance-related traits in groundnut.

Groundnut productivity and quality have been impeded by rising temperatures in semi-arid environments. Hence, understanding the effects and molecular mechanisms of heat stress tolerance will aid in tackling yield losses. In this context, a recombinant inbred line (RIL) population was developed and phenotyped for eight seasons at three locations for agronomic, phenological, and physiological traits under heat stress. A genetic map was constructed using genotyping-by-sequencing with 478 single-nucleotide polymorphism (SNP) loci spanning a map distance of 1,961.39 cM. Quantitative trait locus (QTL) analysis using phenotypic and genotypic data identified 45 major main-effect QTLs for 21 traits. Intriguingly, three QTL clusters (Cluster-1-Ah03, Cluster-2-Ah12, and Cluster-3-Ah20) harbor more than half of the major QTLs (30/45, 66.6%) for various heat tolerant traits, explaining 10.4%-38.6%, 10.6%-44.6%, and 10.1%-49.5% of phenotypic variance, respectively. Furthermore, important candidate genes encoding DHHC-type zinc finger family protein (arahy.J0Y6Y5), peptide transporter 1 (arahy.8ZMT0C), pentatricopeptide repeat-containing protein (arahy.4A4JE9), Ulp1 protease family (arahy.X568GS), Kelch repeat F-box protein (arahy.I7X4PC), FRIGIDA-like protein (arahy.0C3V8Z), and post-illumination chlorophyll fluorescence increase (arahy.92ZGJC) were the underlying three QTL clusters. The putative functions of these genes suggested their involvement in seed development, regulating plant architecture, yield, genesis and growth of plants, flowering time regulation, and photosynthesis. Our results could provide a platform for further fine mapping, gene discovery, and developing markers for genomics-assisted breeding to develop heat-tolerant groundnut varieties.

ZmHOX32 is related to photosynthesis and likely functions in plant architecture of maize.

HOX32, a member of the HD-ZIP III family, functions in the leaf morphogenesis and plant photosynthesis. However, the regulatory mechanism of HOX32 in maize has not been studied and the regulatory relationship in photosynthesis is unclear. We conducted a comprehensive study, including phylogenetic analysis, expression profiling at both transcriptome and translatome levels, subcellular localization, tsCUT&Tag, co-expression analysis, and association analysis with agronomic traits on HOX32 for the dissection of the functional roles of HOX32. ZmHOX32 shows conservation in plants. As expected, maize HOX32 protein is specifically expressed in the nucleus. ZmHOX32 showed constitutively expression at both transcriptome and translatome levels. We uncovered the downstream target genes of ZmHOX32 by tsCUT&Tag and constructed a cascaded regulatory network combining the co-expression networks. Both direct and indirect targets of ZmHOX32 showed significant gene ontology enrichment in terms of photosynthesis in maize. The association study suggested that ZmHOX32 plays an important role in regulation of plant architecture. Our results illustrate a complex regulatory network of HOX32 involving in photosynthesis and plant architecture, which deepens our understanding of the phenotypic variation in plants.

The MAX2-KAI2 module promotes salicylic acid-mediated immune responses in Arabidopsis.

Arabidopsis MORE AXILLARY GROWTH2 (MAX2) is a key component in the strigolactone (SL) and karrikin (KAR) signaling pathways and regulates the degradation of SUPPRESSOR OF MAX2 1/SMAX1-like (SMAX1/SMXL) proteins, which are transcriptional co-repressors that regulate plant architecture, as well as abiotic and biotic stress responses. The max2 mutation reduces resistance against Pseudomonas syringae pv. tomato (Pst). To uncover the mechanism of MAX2-mediated resistance, we evaluated the resistance of various SL and KAR signaling pathway mutants. The resistance of SL-deficient mutants and of dwarf 14 (d14) was similar to that of the wild-type, whereas the resistance of the karrikin insensitive 2 (kai2) mutant was compromised, demonstrating that the KAR signaling pathway, not the SL signaling pathway, positively regulates the immune response. We measured the resistance of smax1 and smxl mutants, as well as the double, triple, and quadruple mutants with max2, which revealed that both the smax1 mutant and smxl6/7/8 triple mutant rescue the low resistance phenotype of max2 and that SMAX1 accumulation diminishes resistance. The susceptibility of smax1D, containing a degradation-insensitive form of SMAX1, further confirmed the SMAX1 function in the resistance. The relationship between the accumulation of SMAX1/SMXLs and disease resistance suggested that the inhibitory activity of SMAX1 to resistance requires SMXL6/7/8. Moreover, the exogenous application of KAR2 enhanced resistance against Pst, but KAR-induced resistance depended on salicylic acid (SA) signaling. Inhibition of karrikin signaling delayed SA-mediated defense responses and inhibited pathogen-induced protein biosynthesis. Together, we propose that the MAX2-KAI2-SMAX1 complex regulates resistance with the assistance of SMXL6/7/8 and SA signaling and that SMAX1/SMXLs possibly form a multimeric complex with their target transcription factors to fine tune immune responses.

QTL Analysis Reveals Conserved and Differential Genetic Regulation of Maize Lateral Angles above the Ear.

Improving the density tolerance and planting density has great importance for increasing maize production. The key to promoting high density planting is breeding maize with a compact canopy architecture, which is mainly influenced by the angles of the leaves and tassel branches above the ear. It is still unclear whether the leaf angles of different stem nodes and tassel branches are controlled by similar genetic regulatory mechanisms, which limits the ability to breed for density-tolerant maize. Here, we developed a population with 571 double haploid lines derived from inbred lines, PHBA6 and Chang7-2, showing significant differences in canopy architecture. Phenotypic and QTL analyses revealed that the genetic regulation mechanism was largely similar for closely adjacent leaves above the ears. In contrast, the regulation mechanisms specifying the angles of distant leaves and the angles of leaves vs. tassel branches are largely different. The liguless1 gene was identified as a candidate gene for QTLs co-regulating the angles of different leaves and the tassel branch, consistent with its known roles in regulating plant architecture. Our findings can be used to develop strategies for the improvement of leaf and tassel architecture through the introduction of trait-specific or pleiotropic genes, thus benefiting the breeding of maize with increased density tolerance in the future.

Improving the adventitious rooting ability of hard-to-root olive (Olea europaea L.) cultivar cuttings through inhibiting strigolactone biosynthesis

Strigolactones (SLs) are synthesized in roots and control plant development. As phytohormones, SLs regulate plant architecture, including roots. Recently, the inhibiting effects of SLs on adventitious rooting have been identified. Olive (Olea europaea L.) is consumed for oil and table in Mediterranean countries and is an economically important crop. Turkey is one of the countries with the highest olive production. Olive has mostly propagated asexually via cuttings, however, the rooting capacities of some agriculturally important olive cultivars are very low. Indole Butyric Acid (IBA) is commonly used to promote the rooting of olive cuttings, however, it can be inadequate. Ayvalık is an easy-to-root cultivar and one of the most common cultivars grown for oil production and Domat is a hard-to-root cultivar in which IBA is insufficient for inducing rooting. In our study, the effects of synthetic SLs rac-GR24 and SLs biosynthesis inhibitor TIS108 on the rooting ability of olive cuttings were investigated. As a result, the adventitious rooting ability was increased when a hard-to-root cultivar was treated with TIS108, indicating a promising future for olive-cutting rooting. Therefore, our study will provide potentially new tools for propagation strategies using SLs in fruit trees.

Understanding the molecular mechanism of leaf morphogenesis in vegetable crops conduces to breeding process.

Leaf morphology can affect the development and yield of plants by regulating plant architecture and photosynthesis. Several factors can determine the final leaf morphology, including the leaf complexity, size, shape, and margin type, which suggests that leaf morphogenesis is a complex regulation network. The formation of diverse leaf morphology is precisely controlled by gene regulation on translation and transcription levels. To further reveal this, more and more genome data has been published for different kinds of vegetable crops and advanced genotyping approaches have also been applied to identify the causal genes for the target traits. Therefore, the studies on the molecular regulation of leaf morphogenesis in vegetable crops have also been largely improved. This review will summarize the progress on identified genes or regulatory mechanisms of leaf morphogenesis and development in vegetable crops. These identified markers can be applied for further molecular-assisted selection (MAS) in vegetable crops. Overall, the review will contribute to understanding the leaf morphology of different crops from the perspective of molecular regulation and shortening the breeding cycle for vegetable crops.

Deciphering the regulatory network of miR156 in plant architecture and abiotic stress resistance of alfalfa (Medicago sativa) by transcriptome sequencing

To improve alfalfa (Medicago sativa) biomass yield and abiotic stress resistance is a challenge due to its autotetraploidy and self-incompatibility. Previously we have generated transgenic alfalfa lines overexpressing miR156 and showing excellent plant architecture. In this study, we investigated the regulation network of miR156 in these transgenic alfalfa lines. Based on 83,202 non-redundant full-length transcripts in a pool of roots, stems, leaves, flowers, pods, and apical buds, we identified 154 candidate genes as potential downstream targets of miR156, especially for M. sativa SQUAMOSA promoter binding protein-like genes (MsSPLs), and then identified 26 MsSPL gene family members in alfalfa genome. The expressions of MsSPL5B and MsSPL3 were downregulated in root and leaf respectively, whereas that of MsSPL4 was significantly downregulated in stem, leaf, root, and apical bud tissues due to miR156 overexpression. We revealed the key role of MsSPL4-mediated regulatory pathways in the substantial correlations between agronomic traits and gene expression, and further found several involved hub genes showing significant correlation with MsSPL4. In addition, MsSPL4 overexpression in tobacco enhanced salinity tolerance but inhibited bud induction. Our results provide strong evidence for understanding the roles of miR156 and MsSPLs in regulating plant architecture and abiotic stress responses in alfalfa.

MiR319 and its target TCP4 involved in plant architecture regulation in Brassica napus

Plant architecture is a collection of genetically controlled crop productivity and adaptation. MicroRNAs (miRNAs) have been proved to function in various biological processes, but little is known about how miRNA regulates plant architecture in rapeseed (Brassica napus L.). In this study, four small RNA libraries and two degradome libraries from shoot apex of normal and rod-like plants were sequenced. A total of 639 miRNA precursors and 16 differentially expressed miRNAs were identified in this study. In addition, 322 targets were identified through degradome sequencing. Among them, 14 targets were further validated via RNA ligase-mediated 5′ rapid amplification of cDNA ends. Transgenic approach showed that increased TCP4 activity in Arabidopsis resulted in premature onset of maturation and reduced plant size along with early flowering and shortened flowering time. miR319-OE lines in Brassica napus exhibited serrated leaves and abnormal development of shoot apical meristem (SAM), which led to the deformed growth of stem and reduced plant height. In conclusion, our study lays the foundation for elucidating miRNA regulate plant architecture and provides new insight into the miR319/TCP4 module regulates plant architecture in rapeseed.

The Effects of High CO2 and Strigolactones on Shoot Branching and Aphid-Plant Compatibility Control in Pea.

Elevated atmospheric CO2 concentrations (eCO2) regulate plant architecture and susceptibility to insects. We explored the mechanisms underpinning these responses in wild type (WT) peas and mutants defective in either strigolactone (SL) synthesis or signaling. All genotypes had increased shoot height and branching, dry weights and carbohydrate levels under eCO2, demonstrating that SLs are not required for shoot acclimation to eCO2. Since shoot levels of jasmonic acid (JA) and salicylic acid (SA) tended to be lower in SL signaling mutants than the WT under ambient conditions, we compared pea aphid performance on these lines under both CO2 conditions. Aphid fecundity was increased in the SL mutants compared to the WT under both ambient and eCO2 conditions. Aphid infestation significantly decreased levels of JA, isopentenyladenine, trans-zeatin and gibberellin A4 and increased ethylene precursor ACC, gibberellin A1, gibberellic acid (GA3) and SA accumulation in all lines. However, GA3 levels were increased less in the SL signaling mutants than the WT. These studies provide new insights into phytohormone responses in this specific aphid/host interaction and suggest that SLs and gibberellins are part of the network of phytohormones that participate in host susceptibility.

Linum usitatissimum FUSCA3–1 regulates plant architecture and seed storage reserve accumulation in Arabidopsis thaliana

Flax ( Linum usitatissimum L.) as a self-pollinated annual diploid crop is grown worldwide primarily for its seed storage reserves of oil and storage proteins. The B3 domain transcription factor AtFUSCA3 (AtFUS3) has been identified as a master regulator of seed storage reserve accumulation in Arabidopsis thaliana . However, the function of LuFUS3 from L. usitatissimum has not yet been assessed. Here, we found that there were two LuFUS3 homologs, LuFUS3–1 and LuFUS3–2, in the L. usitatissimum genome. The subcellular localization and yeast transcriptional activation assays indicated that LuFUS3–1 functions as a transcription factor. Heterogeneous expression of LuFUS3–1 in the A. thaliana wild type background changed the plant architecture, including moderately dwarf stature, wrinkled leaves, increased branch number, altered floral morphology, and shorter and wider siliques, which was at least partially caused by the significantly decreased level of endogenous gibberellins. On the other hand, we demonstrated that LuFUS3–1 enhances the accumulation of seed storage reserves inclusive of oil and storage proteins in A. thaliana seeds. Consistently, LuFUS3–1 promotes the expression of many genes involved in the biosynthesis of seed oil and storage proteins during seed development. These findings provide new insights into the FUS3 function in plant architecture and seed storage reserve accumulation, and also represent a promising target for genetic modification of L. usitatissimum . • LuFUS3-1 functions as a B3 domain transcription factor. • LuFUS3-1 regulates plant architecture at least dependent on gibberellin biosynthesis. • LuFUS3-1 promotes the accumulation of seed oil and storage proteins. • LuFUS3 boosts the expression of genes involved in seed oil and storage protein biosynthesis.

9-cis-β-Apo-10'-carotenal is the precursor of strigolactones in planta.

13C-isotope feeding experiments demonstrate that the apocarotenoid 9-cis-β-apo-10'-carotenal is the precursor of several strigolactones in rice, providing a direct, in planta evidence for its role in strigolactone biosynthesis. Strigolactones (SLs) are plant hormone that regulates plant architecture and mediates rhizospheric communications. Previous in vitro studies using heterogously produced enzymes unraveled the conversion of all-trans-β-carotene via the intermediate 9-cis-β-apo-10'-carotenal into the SL precursor carlactone. However, a direct evidence for the formation of SLs from 9-cis-β-apo-10'-carotenal is still missing. To provide this evidence, we supplied rice seedlings with 13C-labeled 9-cis-β-apo-10'-carotenal and analyzed their SLs by LC-MS. Our results show that 9-cis-β-apo-10'-carotenal is the SL precursor in planta and reveal, for the first time, the application of labeled long-chain apocarotenoids as a promising approach to investigate apocarotenoid metabolism and the genesis of carotenoid-derived growth regulators and signaling molecules.

Integrated genetic mapping and transcriptome analysis reveal the BnaA03.IAA7 protein regulates plant architecture and gibberellin signaling in Brassica napus L.

A novel mutation in the BnaA03.IAA7 protein reduces plant height and enhances gibberellin signaling in Brassica napus L. Rapeseed (Brassica napus) is an excellent and important source for vegetable oil production, but its production is severely affected by lodging. Lodging hinders mechanization and decreases yield, and an ideal solution is semidwarf breeding. Limited by germplasm resources, semidwarf breeding developed slowly in rapeseed. In the current study, a mutant called sdA03 was isolated from EMS-mutagenized lines of Zhongshuang 11 (ZS11). The inheritance analysis showed that phenotypes of sdA03 were controlled by a single semidominant gene. Genetic mapping, RNA-seq and candidate gene analysis identified BnaA03.IAA7 as a candidate gene, and a function test confirmed that the mutated BnaA03.iaa7 regulates plant architecture in a dose-dependent manner. Yeast two-hybrid and transient expression experiments illustrated the P87L substitution in the GWPPV/I degron motif of BnaA03.iaa7 impaired the interaction between BnaA03.IAA7 and TIR1 proteins, and BnaA03.iaa7 prevented ARF from activating the auxin signaling pathway.The gibberellin (GA) content was higher in sdA03 hypocotyls than in those of ZS11. Further expression analysis showed more active gibberellin signaling in hypocotyl and richer expression of GA synthetic genes in root and cotyledon of sdA03 seedlings. Finally, a marker was developed based on the SNP found in BnaA03.iaa7 and used in molecular breeding. The study enriched our understanding of the architectural regulation of rapeseed and provided germplasm resources for breeding.

Melatonin Mediates Axillary Bud Outgrowth by Improving Nitrogen Assimilation and Transport in Rice.

Melatonin plays an important role in plant resistance to biotic and abiotic stresses. However, whether melatonin is involved in the regulation of plant architecture, such as the formation of axillary bud outgrowth or tillering, in rice remains unknown. Here, we found that different concentrations of melatonin influenced axillary bud outgrowth in rice, and moderate melatonin concentrations also alleviated the inhibition of axillary bud outgrowth in the presence of high concentrations of basic amino acids lysine and arginine. Furthermore, transcriptome analysis demonstrated that genes involved in nitrogen metabolism and phytohormone signal transduction pathways may affect axillary bud outgrowth, which is regulated by melatonin. We determined that the differentially expressed genes glutamine synthetase OsGS2 and amino acid transporter OsAAP14, which are involved in nitrogen metabolism and are regulated by melatonin and basic amino acids, were the key regulators of axillary bud outgrowth in rice. In addition, we validated the functions of OsGS2 and OsAAP14 using rice transgenic plants with altered axillary bud outgrowth and tillers. Taken together, these results suggest that melatonin mediates axillary bud outgrowth by improving nitrogen assimilation and transport in rice.

The miR319/TaGAMYB3 module regulates plant architecture and improves grain yield in common wheat (Triticum aestivum).

Plant architecture is a key determinant of crop productivity and adaptation. The highly conserved microRNA319 (miR319) family functions in various biological processes, but little is known about how miR319 regulates plant architecture in wheat (Triticum aestivum). Here, we determined that the miR319/TaGAMYB3 module controls plant architecture and grain yield in common wheat. Repressing tae-miR319 using short tandem target mimics resulted in favorable plant architecture traits, including increased plant height, reduced tiller number, enlarged spikes and flag leaves, and thicker culms, as well as enhanced grain yield in field plot tests. Overexpressing tae-miR319 had the opposite effects on plant architecture and grain yield. Although both TaPCF8 and TaGAMYB3 were identified as miR319 target genes, genetic complementation assays demonstrated that only miR319-resistant TaGAMYB3 (rTaGAMYB3) abolished tae-miR319-mediated growth inhibition of flag leaves and spikes. TaGAMYB3 functions as a transcriptional activator of downstream genes, including TaPSKR1, TaXTH23, TaMADS5 and TaMADS51, by binding to their promoters. Furthermore, TaGAMYB3 physically interacts with TaBA1, an important regulator of spike development, to additively activate the transcription of downstream genes such as TaMADS5. Our findings provide insight into how the miR319/TaGAMYB3 module regulates plant architecture and improves grain yield in common wheat.

Genome-Wide Identification of Gramineae Brassinosteroid-Related Genes and Their Roles in Plant Architecture and Salt Stress Adaptation.

Brassinosteroid-related genes are involved in regulating plant growth and stress responses. However, systematic analysis is limited to Gramineae species, and their roles in plant architecture and salt stress remain unclear. In this study, we identified brassinosteroid-related genes in wheat, barley, maize, and sorghum and investigated their evolutionary relationships, conserved domains, transmembrane topologies, promoter sequences, syntenic relationships, and gene/protein structures. Gene and genome duplications led to considerable differences in gene numbers. Specific domains were revealed in several genes (i.e., HvSPY, HvSMOS1, and ZmLIC), indicating diverse functions. Protein-protein interactions suggested their synergistic functions. Their expression profiles were investigated in wheat and maize, which indicated involvement in adaptation to stress and regulation of plant architecture. Several candidate genes for plant architecture (ZmBZR1 and TaGSK1/2/3/4-3D) and salinity resistance (TaMADS22/47/55-4B, TaGRAS19-4B, and TaBRD1-2A.1) were identified. This study is the first to comprehensively investigate brassinosteroid-related plant architecture genes in four Gramineae species and should help elucidate the biological roles of brassinosteroid-related genes in crops.