Soybean (Glycine max) is a short-day crop whose flowering time is regulated by photoperiod. The long-juvenile trait extends its vegetative phase and increases yield under short-day conditions. Natural variation in J, the major locus controlling this trait, modulates flowering time. We report that the three J-family genes influence soybean flowering time, with the triple mutant Guangzhou Mammoth-2 flowering late under short days by inhibiting transcription of E1-family genes. J-family genes offer promising allelic combinations for breeding.

Sucrose nonfermenting 2 (Snf2) family proteins are the core component of chromatin remodeling complexes that can alter chromatin structure and nucleosome position by utilizing the energy of ATP, playing a vital role in transcription regulation, DNA replication, and DNA damage repair. Snf2 family proteins have been characterized in various species including plants, and they have been found to regulate development and stress responses in Arabidopsis. Soybean (Glycine max) is an important food and economic crop worldwide, unlike other non-leguminous crops, soybeans can form a symbiotic relationship with rhizobia for biological nitrogen fixation. However, little is known about Snf2 family proteins in soybean. In this study, we identified 66 Snf2 family genes in soybean that could be classified into six groups like Arabidopsis, unevenly distributed on 20 soybean chromosomes. Phylogenetic analysis with Arabidopsis revealed that these 66 Snf2 family genes could be divided into 18 subfamilies. Collinear analysis showed that segmental duplication was the main mechanism for expansion of Snf2 genes rather than tandem repeats. Further evolutionary analysis indicated that the duplicated gene pairs had undergone purifying selection. All Snf2 proteins contained seven domains, and each Snf2 protein had at least one SNF2_N domain and one Helicase_C domain. Promoter analysis revealed that most Snf2 genes had cis-elements associated with jasmonic acid, abscisic acid, and nodule specificity in their promoter regions. Microarray data and real-time quantitative PCR (qPCR) analysis revealed that the expression profiles of most Snf2 family genes were detected in both root and nodule tissues, and some of them were found to be significantly downregulated after rhizobial infection. In this study, we conducted a comprehensive analysis of the soybean Snf2 family genes and demonstrated their responsiveness to Rhizobia infection. This provides insight into the potential roles of Snf2 family genes in soybean symbiotic nodulation.

The BYPASS1-related gene (BPS1) encodes a protein with an unknown functional domain that regulates plant organ growth and development by inhibiting the continuous production of a root-derived long-distance signaling molecule called bypass ( bps ). We conducted a comprehensive study to investigate the BPS gene family in soybean and identified twenty-three BPS genes in Glycine max and twenty BPS genes in Glycine soja (wild soybean). Collinearity analysis revealied the existence of multiple orthologs of soybean BPS genes in wild soybean, indicating incomplete conservation between the BPS genes of soybean and wild soybean. Phylogenetic analysis successfully categorized all BPS genes into five distinct groups. We further scrutinized their chromosomal locations, gene structures, conserved motifs, cis-acting elements, and expression patterns. Leveraging publicly available data on genetic variation, phenotypic variation, and single-cell transcriptome sequencing of root nodules, we discovered a potential association between BPS genes and multiple soybean traits, particularly those related to the root nodule phenotype. This pioneering study provides a systematic and comprehensive examination of the BPS gene family in soybean. The findings establish a robust foundation for future investigations into the functional roles of BPS genes in plant growth and development.

In many plants, flowering time is influenced by daylength as an adaptive response. In soybean (Glycine max) cultivars, however, photoperiodic flowering reduces crop yield and quality in high-latitude regions. Under-standing the genetic basis of wild soybean (Glycine soja) adaptation to high latitudes could aid breeding of improved cultivars. Here, we identify the Tof4 (Time of flowering 4) locus, which encodes by an E1-like pro-tein, E1La, that represses flowering and enhances adaptation to high latitudes in wild soybean. Moreover, we found that Tof4 physically associates with the promoters of two important FLOWERING LOCUS T (FT2a and FT5a) and with Tof5 to inhibit their transcription under long photoperiods. The effect of Tof4 on flowering and maturity is mediated by FT2a and FT5a proteins. Intriguingly, Tof4 and the key flowering repressor E1 inde-pendently but additively regulate flowering time, maturity, and grain yield in soybean. We determined that weak alleles of Tof4 have undergone natural selection, facilitating adaptation to high latitudes in wild soybean. Notably, over 71.5% of wild soybean accessions harbor the mutated alleles of Tof4 or a previously reported gain-of-function allele Tof5H2, suggesting that these two loci are the genetic basis of wild soybean adaptation to high latitudes. Almost no cultivated soybean carries the mutated tof4 allele. Introgression of the tof4-1 and Tof5H2 alleles into modern soybean or editing E1 family genes thus represents promising avenues to obtain early-maturity soybean, thereby improving productivity in high latitudes.

Flowering time influences reproductive success in plants and has a significant impact on yield in grain crops. Flowering time is regulated by a variety of environmental factors, with daylength often playing an important role. Crops can be categorized into different types according to their photoperiod requirements for flowering. For instance, long-day crops include wheat ( Triticum aestivum ), barley ( Hordeum vulgare ), and pea ( Pisum sativum ), while short-day crops include rice ( Oryza sativa ), soybean ( Glycine max ), and maize ( Zea mays ). Understanding the molecular regulation of flowering and genotypic variation therein is important for molecular breeding and crop improvement. This paper reviews the regulation of flowering in different crop species with a particular focus on how photoperiod-related genes facilitate adaptation to local environments.

Soybean (Glycine max) is a major source of protein and edible oil world-wide and is cultivated in a wide range of latitudes. However, it is extremely sensitive to photoperiod, which influences flowering time, maturity, and yield, and severely limits soybean latitude adaptation.In this study, a genome-wide association study (GWAS) identified a novel locus in accessions harboring the E1 allele, called Time of flowering 8 (Tof8), which promotes flowering and enhances adaptation to high latitude in cultivated soybean. Gene functional analyses showed that Tof8 is an ortholog of Arabidopsis FKF1.We identified two FKF1 homologs in the soybean genome. Both FKF1 homologs are genetically dependent on E1 by binding to E1 promoter to activate E1 transcription, thus repressing FLOWERING LOCUS T 2a (FT2a) and FT5a transcription, which modulate flowering and maturity through the E1 pathway.We also demonstrate that the natural allele FKF1b(H3) facilitated adaptation of soybean to high-latitude environments and was selected during domestication and improvement, leading to its rapid expansion in cultivated soybean. These findings provide novel insights into the roles of FKF1 in controlling flowering time and maturity in soybean and offer new means to fine-tune adaptation to high latitudes and increase grain yield.

Growth duration is an important agronomic trait that determines the season and area of crop growth. Previous experiments showed that overexpression of nitrate transporter OsNRT2.3b significantly increased rice yield, nitrogen use efficiency, and growth duration. Through screening, we obtained four ethyl methanesulfonate (EMS)-mutagenized mutants with shorter growth duration compared with O8 of OsNRT2.3b overexpression line. The nitrogen translocation efficiency and physiological nitrogen use efficiency of the mutants were not significantly different from O8, which were increased by 24.4% and 14.2%, respectively compared with WT, but the growth duration of the mutant was significantly lower than O8. Analysis of O8 and mutants showed that the growth duration positively correlated with grain weight per panicle, grain yield, and nitrogen recovery efficiency. In conclusion, our results provide a new idea for balancing rice yield and growth duration.

The intrinsic relationship between biscuit quality and starch physicochemical properties was studied using a set of wheat lines with HMW-GS absence of Ningmai 9. The results showed that, compared with wild type, wheat lines with HMW-GS absence had higher amylose content, volume proportion of A-type starch granule, relative crystallinity (RC), peak and final viscosities, gelatinization peak temperature (Tp) and conclusion temperature (Tc), and consistency coefficient (K) of flour paste. Conversely, lower amylopectin content and ratio of absorbance 1047/1022 cm-1 (IR2) were found in these wheat lines. As a result, biscuits made of wheat lines with HMW-GS absence had better quality with higher spread ratio and lower hardness. Pearson correlation analysis showed that the difference between minor compounds contents and biscuit quality was not significant, while amylose content, volume proportion of A-type starch granule, RC, and K value were positively correlated with biscuit quality, confirming significant effects of starch physicochemical properties on biscuit quality and effective indicators for improvement of biscuit quality. Molecular mechanisms were proposed for all these observations, which not only shed light on how physicochemical properties of starch affects biscuit quality but also identified high-quality soft wheat for biscuit making by selecting starch with appropriate physicochemical features.

Excessive nitrogen fertiliser use reduces nitrogen use efficiency and causes significant damage to the environment. Carbon fertilisers have the advantage of improving soil fertility; however, the effects of carbon and nitrogen fertilisers on rice yield and quality are not clear. In this study, the nitrogen-efficient line (OsNRT2.3b-overexpressing [O8]) and wild type (WT) were treated with different levels of nitrogen and carbon fertilisers under field conditions to study the effects of different fertilisation treatments on rice quality. The results showed that the appearance, nutrition, and taste qualities of O8 were generally high compared with WT under various fertilisation treatment conditions in 2019 and 2020. Compared with 90 kg/ha and 270 kg/ha nitrogen fertiliser, a single application of 90 kg/ha and 270 kg/ha carbon fertiliser significantly reduced the protein content of O8 by approximately 37.08% and 35.50% in 2019 and 2020, respectively, compared with WT, and improved the eating quality of O8 and WT. However, the replacement of nitrogen fertiliser with 20% carbon fertiliser did not improve the eating quality of O8 and WT compared with a single application of nitrogen fertiliser. This study identifies a high-quality gene, OsNRT2.3b, for breeding high-quality rice and provides a theoretical basis for obtaining high-quality rice and molecular breeding.

Vesicular trafficking plays critical roles in cell expansion in yeast and mammals, but information linking vesicular trafficking and cell expansion in plants is limited. Here, we isolated and characterized a rice (Oryza sativa) mutant, decreased plant height 1-1 (dph1-1), which exhibited a wide spectrum of developmental phenotypes, including reduced plant height and smaller panicles and grains. Cytological analysis revealed that limited cell expansion was responsible for the dph1-1 mutant phenotype compared to the wild-type. Map-based cloning revealed that DPH1 encodes a plant-specific protein, OsSCD2, which is homologous to Arabidopsis (Arabidopsis thaliana) STOMATAL CYTOKINESIS DEFECTIVE2 (SCD2). Subcellular localization revealed that OsSCD2 is associated with clathrin. Confocal microscopy showed that the dph1-1 mutant has defective endocytosis and post-Golgi trafficking. Biochemical and confocal data indicated that OsSCD2 physically interacts with OsSCD1 and that they are associated with intracellular structures that colocalize with microtubules. Furthermore, we found that cellulose synthesis was affected in the dph1-1 mutant, evidenced by reduced cellulose synthase gene accumulation at the transcript and protein levels, most likely resulting from an impaired localization pattern. Our results suggest that OsSCD2 is involved in clathrin-related vesicular trafficking with an important role in maintaining plant growth in rice.

Bread wheat is a staple food crop that is consumed worldwide. In this study, using widely targeted LC-MS/MS, we conducted a high-throughput metabolomic analysis and determined the contents and spatial distribution of metabolites in pearled fractions of the dried kernels of six representative bread wheat varieties cultivated in China. Our aim was to explore the cultivars and pearling fractions with a view toward developing functional food products. We accordingly identified notable differences in the nutrient and bioactive metabolomes, and established that the pearling fractions of each cultivar had distinct metabolic profiles. Flavonoids varied the most amongst the cultivars and were found in higher concentration in the outer layers of the grain, but only at low concentrations in the kernel. Data from this study add further evidence of benefits of whole grain wheat consumption but, specifically, medium-gluten and pigmented wheat offer other nutrient and bioactive benefits whole grain products.

Soybean (Glycine max) grows in a wide range of latitudes, but it is extremely sensitive to photoperiod, which reduces its yield and ability to adapt to different environments. Therefore, understanding of the genetic basis of soybean adaptation is of great significance for breeding and improvement. Here, we characterized Tof18 (SOC1a) that conditions early flowering and growth habit under both short-day and long-day conditions. Molecular analysis confirmed that the two SOC1 homologs present in soybeans (SOC1a and SOC1b) underwent evolutionary functional divergence, with SOC1a having stronger effects on flowering time and stem node number than SOC1b due to transcriptional differences. soc1a soc1b double mutants showed stronger functional effects than either of the single mutants, perhaps due to the formation of SOC1a and SOC1b homodimers or heterodimers. Additionally, Tof18/SOC1a improves the latitudinal adaptation of cultivated soybeans, highlighting the functional importance of SOC1a. The Tof18G allele facilitates adaptation to high latitudes, whereas Tof18A facilitates adaptation to low latitudes. We demonstrated that SOC1s contribute to floral induction in both leaves and shoot apex through inter-regulation with FTs. The SOC1a-SOC1b-Dt2 complex plays essential roles in stem growth habit by directly binding to the regulatory sequence of Dt1, making the genes encoding these proteins potential targets for genome editing to improve soybean yield via molecular breeding. Since the natural Tof18A allele increases node number, introgressing this allele into modern cultivars could improve yields, which would help optimize land use for food production in the face of population growth and global warming.

Protein storage vacuoles (PSVs) are unique organelles that accumulate storage proteins in plant seeds. Although morphological evidence points to the existence of multiple PSV-trafficking pathways for storage protein targeting, the molecular mechanisms that regulate these processes remain mostly unknown. Here, we report the functional characterization of the rice (Oryza sativa) glutelin precursor accumulation7 (gpa7) mutant, which over-accumulates 57-kDa glutelin precursors in dry seeds. Cytological and immunocytochemistry studies revealed that the gpa7 mutant exhibits abnormal accumulation of storage prevacuolar compartment-like structures, accompanied by the partial mistargeting of glutelins to the extracellular space. The gpa7 mutant was altered in the CCZ1 locus, which encodes the rice homolog of Arabidopsis (Arabidopsis thaliana) CALCIUM CAFFEINE ZINC SENSITIVITY1a (CCZ1a) and CCZ1b. Biochemical evidence showed that rice CCZ1 interacts with MONENSIN SENSITIVITY1 (MON1) and that these proteins function together as the Rat brain 5 (Rab5) effector and the Rab7 guanine nucleotide exchange factor (GEF). Notably, loss of CCZ1 function promoted the endosomal localization of vacuolar protein sorting-associated protein 9 (VPS9), which is the GEF for Rab5 in plants. Together, our results indicate that the MON1-CCZ1 complex is involved in post-Golgi trafficking of rice storage protein through a Rab5- and Rab7-dependent pathway.

S genes are the key genes that cause plant self-incompatibility, to find out the key S genes and understand molecular mechanism of self-incompatibility in chrysanthemum, the stigmas and anthers at different developmental stages of 'Q10-22-2'—a self-incompatible chrysanthemum cultivar, were used for RNA sequencing. After bioinformatics analysis, 13 candidate pistil S genes and five candidate pollen S genes were excavated. A potential pistil S gene was cloned and named as CmSRK1. Meanwhile, a potential pollen S gene was cloned and named as CmPCP1. qRT-PCR revealed that CmSRK1 was specifically expressed in mature stigmas, and CmPCP1 was specifically expressed in anthers 3 d before maturation. Subcellular localization showed that both CmSRK1 and CmPCP1 were located in the nucleus and the cell membrane. Transcriptional activation activity analysis indicated that both of the two proteins had no transcriptional activation activity. Yeast two hybrid assay showed that there was no interaction between CmSRK1 and CmPCP1. CmSRK1 was constructed on the expression vector containing stigma-specific promoter, and CmPCP1 was constructed on the expression vector containing pollen-specific promoter, they were then transformed into Arabidopsis thaliana. Artificial hybridization was performed with transgenic lines containing CmSRK1 as the female parents, and transgenic lines containing CmPCP1 as the male parents. The hybridization results showed that seed sets of two transgenic lines were 19.62% and 11.64%, respectively, while cross-pollinated seed sets of Col-0 was 84.43%. Therefore, it was speculated that CmSRK1 and CmPCP1 might be pistil and pollen S genes of chrysanthemum, respectively, and SI of chrysanthemum belonged to SSI.

Chrysanthemum (Chrysanthemum x morifolium) is an important horticultural crop with great ornamental and economic value. Most chrysanthemum cultivars are self-incompatible, which limits the creation of pure chrysanthemum lines which are the material basis for breeding and theoretical research. Here, artificial self-pollination and natural cross-pollination were used to analyze the self-compatibility (SC) and cross-compatibility (CC) of 85 chrysanthemums cultivated by our research group. By conducting a phenotypic study, observing of pollen germination in vitro and in vivo and gene expression analyses, we explored the reasons for the self-compatibility of chrysanthemum and identified self-compatible mutants. It was found that there were wide variations in self-compatibility among different chrysanthemum cultivars and the SC index ranged from 0 to 5.64, which could classify the chrysanthemum cultivars into three types: self-compatibility (SC index >= 1), self-incompatibility (SI, 1 > SC index > 0) and high self-incompatibility (SC index = 0). Seven chrysanthemum cultivars with self-pollinated progenies ('Hongguan', K183 8, A22, B12, D87, D76 and S8) were selected, and the pollen germination rates in vitro and in vivo were not significantly related to SC. The pollen tube of `Hongguan' could pass through its own stigma and grow toward the embryo sac along the style, and there was no obvious inbreeding depression in self-pollinated progenies of `Hongguan'. In addition, we found that one S-locus receptor kinase (SRK), two S-locus cysteine-rich protein (SCR), two exocyst complex component EXo70A1 (Exo70A1) and one thioredoxin h-like 1 (THL1) were important for regulating self-compatibility of C. x morifolium. Our results not only laid a material foundation for creating pure lines through continuous self-pollination and analyzing the regulation mechanism underlying SC variation but also provided the possibility for theoretical research on the important ornamental traits of chrysanthemums in the future.

KEY MESSAGE:OsVIN2, a vacuolar invertase, affects grain size and yield by altering sugar composition, transport, and starch accumulation in rice. Grain size, a major determinant of rice yield, is influenced by many developmental and environmental factors. Sugar metabolism plays vital roles in plant development. However, the way in which sugar metabolism affects rice grain size remains largely elusive. In this study, we characterized the small grain-size rice mutant sgs1. Histological analyses showed that reduced spikelet hull and endosperm size results from decreased cell size rather than cell number. Map-based cloning and complementation tests revealed that a DaiZ7 transposon insertion in a vacuolar invertase gene OsVIN2 is responsible for the mutant phenotype. Subcellular distribution and biochemical analysis indicated that OsVIN2 is located in the vacuolar lumen, and that its sucrose hydrolysis activity is maintained under acidic conditions. Furthermore, an altered sugar content with increased sucrose and decreased hexose levels, as well as changes in invertase and sucrose synthase activities, sugar transport gene expression, and starch constitution in sgs1 implies that OsVIN2 affects sucrose metabolism, including sugar composition, transport, and conversion from the source to the sink organs. Collectively, OsVIN2 is involved in sugar metabolism, and thus regulates grain size; our findings provide insights into grain development and also suggest a potential strategy to improve grain quality and yield in rice.

Starch is a major storage substance in cereal grains, and starch biosynthesis is a complex process. In order to elucidate regulation of the starch biosynthesis pathway, we screened a series of rice (Oryza sativa L.) endosperm mutants. In this study, we identified a floury white-core endosperm mutant named floury endosperm12 (flo12). The flo12 mutant exhibited loosely packed starch granules and a lower thousand kernel weight compared to wild type. Semithin sections revealed that compound starch grains (SG) in flo12 interior endosperm cells were developed abnormally. Furthermore, amylose content was decreased, while total protein content was significantly increased in flo12 grains. Map-based cloning showed that FLO12 encodes rice alanine aminotransferase 1 (OsAlaAT1). OsAlaAT1 is highly expressed in developing endosperm. Subcellular localization showed that OsAlaAT1 is localized in the cytosol. Moreover, the expression of most starch synthesis-related genes was decreased, while most of the storage protein coding genes had elevated expression levels in the flo12 mutant. In addition, overexpression of the OsAlaAT1 gene increased grain weight. In brief, we demonstrated that OsAlaAT1 regulates carbon and nitrogen metabolism, which provides a new insight for the improvement of rice quality and yield.

81 SNPs were identified for three inflorescence-related traits, in which 15 were highly favorable. Two dCAPS markers were developed for future MAS breeding, and six candidate genes were predicted. Chrysanthemum is a leading ornamental species worldwide and demonstrates a wealth of morphological variation. Knowledge about the genetic basis of its phenotypic variation for key horticultural traits can contribute to its effective management and genetic improvement. In this study, we conducted a genome-wide association study (GWAS) based on two years of phenotype data and a set of 92,617 single nucleotide polymorphisms (SNPs) using a panel of 107 diverse cut chrysanthemums to dissect the genetic control of three inflorescence-related traits. A total of 81 SNPs were significantly associated with the three inflorescence-related traits (capitulum diameter, number of ray florets and flowering time) in at least one environment, with an individual allele explaining 22.72–38.67% of the phenotypic variation. Fifteen highly favorable alleles were identified for the three target traits by computing the phenotypic effect values for the stable associations detected in 2 year-long trials at each locus. Dosage pyramiding effects of the highly favorable SNP alleles and significant linear correlations between highly favorable allele numbers and corresponding phenotypic performance were observed. Two highly favorable SNP alleles correlating to flowering time and capitulum diameter were converted to derived cleaved amplified polymorphic sequence (dCAPS) markers to facilitate future breeding. Finally, six putative candidate genes were identified that contribute to flowering time and capitulum diameter. These results serve as a foundation for analyzing the genetic mechanisms underlying important horticultural traits and provide valuable insights into molecular marker-assisted selection (MAS) in chrysanthemum breeding programs.

Ubiquitination and deubiquitination are reversible processes that play crucial roles in regulating organ size in plants. However, information linking deubiquitination and seed size in rice (Oryza sativa) is limited. Here, we characterized a dominant large-grain mutant, large grain1-D (lg1-D), with a 30.8% increase in seed width and a 34.5% increase in 1,000-grain weight relative to the wild type. The lg1-D mutant had more cells oriented in the lateral direction of the spikelet hull compared with the wild type. Map-based cloning showed that LG1 encodes a constitutively expressed ubiquitin-specific protease15 (OsUBP15) that possesses deubiquitination activity in vitro. Loss-of-function and down-regulated expression of OsUBP15 produced narrower and smaller grains than the control. A set of in vivo experiments indicated that the mutant Osubp15 had enhanced protein stability relative to wild-type OsUBP15. Further experiments verified that OsDA1 directly interacted with OsUBP15. Genetic data indicated that OsUBP15 and GRAIN WIDTH 2 (GW2) were not independent in regulating grain width and size. In summary, we identified OsUBP15 as a positive regulator of grain width and size in rice and provide a promising strategy for improvement of grain yield by pyramiding OsUBP15 and gw2.

The transient elevation of cytoplasmic calcium is essential for pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI). However, the calcium channels responsible for this process have remained unknown. Here, we show that rice CDS1 (CELL DEATH and SUSCEPTIBLE to BLAST 1) encoding OsCNGC9, a cyclic nucleotide-gated channel protein, positively regulates the resistance to rice blast disease. We show that OsCNGC9 mediates PAMP-induced Ca2+ influx and that this event is critical for PAMPs-triggered ROS burst and induction of PTI-related defense gene expression. We further show that a PTI-related receptor-like cytoplasmic kinase OsRLCK185 physically interacts with and phosphorylates OsCNGC9 to activate its channel activity. Our results suggest a signaling cascade linking pattern recognition to calcium channel activation, which is required for initiation of PTI and disease resistance in rice.