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SUMMARY: Grain weight, grain number per panicle, and the number of panicles are the three factors that determine rice (Oryza sativa L.) yield. Of these, grain weight, which not only directly determines rice yield but also influences appearance and quality, is often considered the most important for rice production. Here, we describe OsNF‐YC1, a member of the NF‐Y transcription factor family that regulates rice grain size. OsNF‐YC1 knockout plants (osnf‐yc1), obtained using CRISPR‐Cas9 technology, showed reduced grain weight due to reduced width and thickness, with no change in grain length, leading to a slenderer grain shape. Downregulation of OsNF‐YC1 using RNA interference resulted in similar grain phenotypes as osnf‐yc1. OsNF‐YC1 affects grain formation by regulating both cell proliferation and cell expansion. OsNF‐YC1 localizes in both the nucleus and cytoplasm, has transcriptional activation activity at both the N‐terminus and C‐terminus, and is highly expressed in young panicles. OsNF‐YC1 interacts with OsMADS1 both in vivo and in vitro. Further analysis showed that the histone‐like structural CBFD‐NFYB‐HMF domain of OsNF‐YC1 conserved in the OsNF‐YC transcription factor family can directly interact with the MADS‐box domain of OsMADS1 to enhance its transcriptional activation activity. This interaction positively regulates the expression of OsMADS55, the direct downstream target of OsMADS1. Therefore, this paper reveals a potential grain size regulation pathway controlled by an OsNF‐YC1‐OsMADS1‐OsMADS55 module in rice. Significance Statement: This paper described the molecular mechanism of OsNF‐YC1 regulating rice grain size formation, a potential pathway of OsNF‐YC1‐OsMADS1‐OsMADS55. OsNF‐YC1 knockout plants displayed the slender grain size with the narrower grain width than the controls, the yield of which was not changed due to the number of grains per panicle of the osnf‐yc1 mutant is increased. Therefore, OsNF‐YC1 can be used for improving rice appearance quality and yield, which has a potential value in practical application. [ABSTRACT FROM AUTHOR]

In situ conserved wild rice (Oryza rufipogon Griff.) is a promising source of alleles for improving rice production worldwide. In this study, we conducted a genomic analysis of an in situ conserved wild rice population (Guiping wild rice) growing at the center of wild rice genetic diversity in South China. Differences in the plant architecture in this population were investigated. An analysis using molecular markers revealed the substantial genetic diversity in this population, which was divided into subgroups according to the plant architecture. After resequencing representative individuals, the Guiping wild rice population was compared with other O. rufipogon and Oryza sativa populations. The results indicated that this in situ conserved wild rice population has a unique genetic structure, with genes that were introgressed from aromatic and O. sativa ssp. indica and japonica populations. The QTLs associated with plant architecture in this population were detected via a pair-wise comparison analysis of the sequencing data for multiple DNA pools. The results suggested that a heading date-related gene (DHD1) might be associated with variations in plant architecture and may have originated in cultivated rice. Our findings provide researchers with useful insights for future genomic analyses of in situ conserved wild rice populations. [ABSTRACT FROM AUTHOR]

Inorganic phosphate (Pi) is taken up by plant roots and translocated via phosphate transporters. Previously, we showed that phosphate transporter OsPHT1;4 in the PHT1 family participates in phosphate acquisition and mobilization; it facilitates the embryo development of Japonica rice Nipponbare. This study investigated the potential of manipulating the expression of OsPHT1;4 to increase Pi acquisition efficiency and crop productivity in rice cv. Wuyunjing 7 (WYJ 7), a cultivar widely grown in Yangtze River Delta of China. The OsPHT1;4 overexpression lines and wild-type WYJ 7 were treated under different Pi conditions in hydroponic and field experiments. Quantitative real-time RT-PCR analysis and the transgenic plants expressing GUS reporter gene indicate strong expression of OsPHT1;4 in roots and leaf collars of cv. WYJ 7. The total P contents in shoots of the OsPHT1;4-overexpressing plants were significantly higher under Pi-deficient hydroponic conditions than the wild type under Pi sufficiency and deficiency. 33Pi uptake and translocation assays confirmed the results. In the field condition, OsPHT1;4 overexpression lines had a higher P concentration in tissues than the wild type control, and the panicle performance of the overexpression lines including the grain yield was improved as well. Taken together, our results show that OsPHT1;4 plays an important role in the acquisition and mobilization of Pi in WYJ 7, especially under Pi deficiency. The study highlights the importance of OsPHT1;4 in improving the agronomic traits of the widely grown rice cultivar in China. [ABSTRACT FROM AUTHOR]

Moderate curling generally causes upright leaf blades, which favors the establishment of ideal plant architecture and increases the photosynthetic efficiency of the population, both of which are desirable traits for super hybrid rice (Oryza sativa L.). In this study, we identified a novel curled-leaf mutant, curled flag leaf 2 (cfl2), which shows specific curling at the base of the flag leaf owing to abnormal epidermal development, caused by enlarged bulliform cells and increased number of papillae with the disordered distribution. Map-based cloning reveals that CFL2 encodes a cytochrome P450 protein and corresponds to the previously reported OsCYP96B4. CFL2 was expressed in all analyzed tissues with differential abundance and was downregulated in the clf1 mutant [a mutant harbors a mutation in the homeodomain leucine zipper IV (HD-ZIP IV) transcription factor Roc5 ]. Yeast one-hybrid and transient expression assays confirm that Roc5 could directly bind to the cis -element L1 box in the promoter of CFL2 before activating CFL2 expression. RNA sequencing reveals that genes associated with cellulose biosynthesis and cell wall-related processes were significantly upregulated in the cfl2 mutant. The components of cell wall, such as lignin, cellulose, and some kinds of monosaccharide, were altered dramatically in the cfl2 mutant when compared with wild-type "Jinhui10" (WT). Taken together, CFL2 , as a target gene of Roc5 , plays an important role in the regulation of flag leaf shape by influencing epidermis and cell wall development. [ABSTRACT FROM AUTHOR]

Members of the Low Phosphate Root (LPR) family have been identified in rice (Oryza sativa) and expression analyses have been conducted. Here, we investigated the functions of one of the five members in rice, LPR5. qRT-PCR and promoter– GUS reporter analyses indicated that under Pi-sufficient conditions OsLPR5 was highly expressed in the roots, and specific expression occurred in the leaf collars and nodes, and its expression was increased under Pi-deficient conditions. In vitro analysis of the purified OsLPR5 protein showed that it exhibited ferroxidase activity. Overexpression of OsLPR5 triggered higher ferroxidase activity, and elevated concentrations of Fe(III) in the xylem sap and of total Fe in the roots and shoots. Transient expression of OsLPR5 in Nicotiana benthamiana provided evidence of its subcellular localization to the cell wall and endoplasmic reticulum. Knockout mutation in OsLPR5 by means of CRISPR-Cas9 resulted in adverse effects on Pi translocation, on the relative expression of Cis-NAT OsPHO1;2 , and on several morphological traits, including root development and yield potential. Our results indicate that ferroxidase-dependent OsLPR5 has both a broad-spectrum influence on growth and development in rice as well as affecting a subset of physiological and molecular traits that govern Pi homeostasis. [ABSTRACT FROM AUTHOR]

Sucrose-proton symporters play important roles in carbohydrate transport during plant growth and development. Their physiological functions have only been partly characterized and their regulation mechanism is largely unclear. Here we report that the knockout of a sucrose transporter gene, OsSUT1, by CRISPR-Cas9 mediated gene editing resulted in a slightly dwarf size and complete infertility of the gene's homozygous mutants. Observation of caryopsis development revealed that the endosperm of OsSUT1 mutants failed to cellularize and did not show any sign of seed-filling. Consistently, OsSUT1 was identified to express strongly in developing caryopsis of wild-type rice, particularly in the nucellar epidermis and aleurone which are critical for the uptake of nutrients into the endosperm. These results indicate that OsSUT1 is indispensable during the rice reproductive stage particularly for caryopsis development. Interestingly, OsSUT1 possesses at least 6 alternative splicing transcripts, including the 4 transcripts deposited previously and the other two identified by us. The differences among these transcripts primarily lie in their coding region of the 3′ end and 3′ UTR region. Real-time PCR showed that 4 of the 6 transcripts had different expressional patterns during rice vegetative and reproductive growth stages. Given the versatility of the gene, addressing its alternative splicing mechanism may expand our understanding of SUT's function substantially. [ABSTRACT FROM AUTHOR]

The improvement of rice leaf morphology is an important goal of rice plant breeding. Studying its molecular development mechanism will facilitate the breeding of a maximally efficient plant type. To further study the developmental mechanism of rice leaves, a dwarf broad-leaf mutant, g44, was obtained by ethyl methanesulfonate mutagenesis of the rice restorer line Jinhui10, and genetic analysis indicated that the trait was controlled by a pair of recessive nuclear gene. The locus was fine mapped on chromosome 6 with 16-kb physical distance. Sequencing revealed that an A-T base substitution occurred in the mutant, resulting in the early termination of LOC_Os06g03710/DLT translation in g44 and yielding only one residual peptide containing 558 amino acids. The phenotypic and sequencing results of the complementary transgenic plants and the allelic mutant g44-1 further confirmed this result. Mutant plants grown under field conditions showed dwarfing, a leaf colour of dark green, leaf widening, and grain enlargement. Paraffin section and cryosection analysis showed that the increase in the number of large vascular bundles and small vascular bundles and the increase in the spacing between adjacent small vascular bundles were the main reasons for the broadening of mutant leaves. In addition, the increase in thickness of the mesophyll cell layer may be the main reason for the increase in leaf colour darkness and net photosynthetic rate. The qPCR results preliminarily predicted that the increase in the number of mutant cells was caused by an enhanced cell division ability, and the DLT/G44 gene was proposed to participate in the regulation of cell division by suppressing the expression of cyclin-related genes. [ABSTRACT FROM AUTHOR]

The mechanism underlying the ability of rice to germinate underwater is a largely enigmatic but key research question highly relevant to rice cultivation. Moreover, although rice is known to accumulate salicylic acid (SA), SA biosynthesis is poorly defined, and its role in underwater germination is unknown. It is also unclear whether peroxisomes, organelles essential to oilseed germination and rice SA accumulation, play a role in rice germination. Here, we show that submerged imbibition of rice seeds induces SA accumulation to promote germination in submergence. Two submergence-induced peroxisomal Oryza sativa cinnamate:CoA ligases (OsCNLs) are required for this SA accumulation. SA exerts this germination-promoting function by inducing indole-acetic acid (IAA) catabolism through the IAA-amino acid conjugating enzyme GH3. The metabolic cascade we identified may potentially be adopted in agriculture to improve the underwater germination of submergence-intolerant rice varieties. SA pretreatment is also a promising strategy to improve submerged rice germination in the field. [Display omitted] • OsCNLs are induced in rice submerged imbibition to synthesize salicylic acid (SA) • Peroxisomal OsCNLs are key enzymes in the SA biosynthetic pathway in rice • SA promotes submerged germination via OsGH3-mediated indole-acetic acid catabolism • SA pretreatment is a cost-effective strategy to improve rice submerged germination Wang et al. demonstrated a peroxisomal cinnamate:CoA ligase (CNL)-dependent phytohormone metabolic cascade in rice. Submerged imbibition-induced salicylic acid (SA) biosynthesis promotes anaerobic germination by inducing OsGH3-mediated indole-acetic acid (IAA)-amino acid conjugation, thus releasing IAA's inhibition of germination under water submergence conditions. [ABSTRACT FROM AUTHOR]

As members of the pathogenesis-related protein (PR)-2 family, β-1,3-glucanases play pivotal roles in plant defense. Previous study showed that the rice genome contains 16 genes encoding putative β-1,3-glucanases, and the β-1,3-glucanases in subfamily A were deduced to be involved in plant defense. However, there was limited direct evidence. In this study, the expression of rice β-1,3-glucanases Gns2–Gns6 belonging to subfamily A in rice plant infection with Magnaporthe oryzae was investigated, and the enhanced expression of Gns6 during infection confirmed its crucial role in the defense of rice seedlings. Enzymological characterization revealed that Gns6 preferentially hydrolyzed laminarin, pachymaran, and yeast glucan. The β-1,3; 1,6-glucanase Gns6 exhibited a specific activity of 1.2 U/mg with laminarin as the substrate. In addition, Gns6 could hydrolyze laminarin via an endo-type mechanism, yielding a series of oligosaccharides with various degrees of polymerization that are known immune elicitors in plants. Moreover, Gns6 exhibited a significant inhibitory effect against the formation of the germ tubes and appressoria, with potential applications in plant protection. Taken together, this study shows that Gns6 is an essential effector in the defensive response of rice against pathogenic fungi. [ABSTRACT FROM AUTHOR]

Abstract Small ubiquitin-related modifier (SUMO) is a post-translational modification of proteins that has important roles in plant growth and development as well as nutrition study. OsSIZ1, a SUMO E3 ligase in rice (Oryza sativa), exerts regulatory influence on nitrogen (N) homeostasis. Here, we investigated the biological function of OsSIZ2 , a paralog of OsSIZ1 , in the responses to nitrogen, anther dehiscence, and seed length using a reverse genetics approach. The expression of OsSIZ2 was increased during N deficiency. Under -N condition, total N concentration in the root of OsSIZ2 -Ri plants and ossiz2 was significantly increased compared with wild type. Further, 15N-labelled uptake assay revealed the role of OsSIZ2 in acquisition and mobilization of N. Moreover, qRT-PCR analyses revealed that several genes involved in the maintenance of N homeostasis were altered in OsSIZ2 mutants. In addition, ossiz2 indicated obvious defects in anther dehiscence, pollen fertility, and seed set percentage. Interestingly, however, the seed length was longer in the mutant compared with wild type. Overall, these results suggest pivotal roles of OsSIZ2 in regulating homeostasis of N and different agronomic traits including anther and seed development. [ABSTRACT FROM AUTHOR]

SUMOylation is a post-translational modification process that comprises a tandem enzymatic cascade, i.e., maturation, activation, conjugation, and ligation of a small ubiquitin-like modifier, which triggers the modulated activities and transport of the cellular proteins to other areas of the cell. In Oryza sativa (rice), OsSIZ1/2 encoding E3 SUMO ligase exerts regulatory influences on Pi homeostasis and developmental responses. However, the role of OsSAE1a , SUMO E1 activating enzyme, in regulating phosphate (Pi) utilization and/or growth and development is not known in rice and was thus investigated in this study. The qRT-PCR assay revealed a constitutive and variable spatiotemporal expression pattern of OsSAE1a in the vegetative and reproductive tissues and was comparable in the root and shoot grown under different Pi regimes. RNAi-mediated suppression of OsSAE1a exerted variable effects on the concentrations of Pi and total P in different tissues, uptake and distribution of 32Pi, and relative expression levels of several genes that play pivotal roles in the maintenance of Pi homeostasis. The effects of the mutation in OsSAE1a were also evident in the vegetative and reproductive traits of rice during growth in a hydroponic system and pot soil, respectively. Overall, these results suggest a broad-spectrum role of OsSAE1a in the maintenance of Pi homeostasis and regulating growth and development. [ABSTRACT FROM AUTHOR]