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The morphological development of rice (Oryza sativa L.) leaves is closely related to plant architecture, physiological activities, and resistance. However, it is unclear whether there is a co-regulatory relationship between the morphological development of leaves and adaptation to drought environment. In this study, a drought-sensitive, roll-enhanced, and narrow-leaf mutant (renl1) was induced from a semi-rolled leaf mutant (srl1) by ethyl methane sulfonate (EMS), which was obtained from Nipponbare (NPB) through EMS. Map-based cloning and functional validation showed that RENL1 encodes a cellulose synthase, allelic to NRL1/OsCLSD4. The RENL1 mutation resulted in reduced vascular bundles, vesicular cells, cellulose, and hemicellulose contents in cell walls, diminishing the water-holding capacity of leaves. In addition, the root system of the renl1 mutant was poorly developed and its ability to scavenge reactive oxygen species (ROS) was decreased, leading to an increase in ROS after drought stress. Meanwhile, genetic results showed that RENL1 and SRL1 synergistically regulated cell wall components. Our results revealed a theoretical basis for further elucidating the molecular regulation mechanism of cellulose on rice drought tolerance, and provided a new genetic resource for enhancing the synergistic regulation network of plant type and stress resistance, thereby realizing simultaneous improvement of multiple traits in rice.

A study published in the Journal of Integrative Plant Biology examines the role of carotenoid isomerase in regulating rice tillering and grain productivity. The researchers found that carotenoid isomerase activity and content are crucial for maintaining the appropriate number of tillers, photosynthesis, and grain yield. They also discovered that interactions between the strigolactone and abscisic acid pathways contribute to regulating tiller formation. The study identifies a gene called DUONIE (DN) that controls tillering and grain size through the carotenoid biosynthesis pathway. Mutations in the DN gene result in more tiller buds and faster elongation, leading to more tillers but smaller grains. This research provides valuable insights into the genetic mechanisms that influence rice architecture and grain productivity. [Extracted from the article]

Drought stress is a major environmental factor that limits the growth, development, and yield of rice (Oryza sativa L.). Histone deacetylases (HDACs) are involved in the regulation of drought stress responses. HDA704 is an RPD3/HDA1 class HDAC that mediates the deacetylation of H4K8 (lysine 8 of histone H4) for drought tolerance in rice. In this study, we show that plants overexpressing HDA704 (HDA704‐OE) are resistant to drought stress and sensitive to abscisic acid (ABA), whereas HDA704 knockout mutant (hda704) plants displayed decreased drought tolerance and ABA sensitivity. Transcriptome analysis revealed that HDA704 regulates the expression of ABA‐related genes in response to drought stress. Moreover, HDA704 was recruited by a drought‐resistant transcription factor, WAX SYNTHESIS REGULATORY 2 (OsWR2), and co‐regulated the expression of the ABA biosynthesis genes NINE‐CIS‐EPOXYCAROTENOID DIOXYGENASE 3 (NCED3), NCED4, and NCED5 under drought stress. HDA704 also repressed the expression of ABA‐INSENSITIVE 5 (OsABI5) and DWARF AND SMALL SEED 1 (OsDSS1) by regulating H4K8ac levels in the promoter regions in response to polyethylene glycol 6000 treatment. In agreement, the loss of OsABI5 function increased resistance to dehydration stress in rice. Our results demonstrate that HDA704 is a positive regulator of the drought stress response and offers avenues for improving drought resistance in rice. [ABSTRACT FROM AUTHOR]

Summary: Plant architecture and stress tolerance play important roles in rice breeding. Specific leaf morphologies and ideal plant architecture can effectively improve both abiotic stress resistance and rice grain yield. However, the mechanism by which plants simultaneously regulate leaf morphogenesis and stress resistance remains elusive. Here, we report that SRL10, which encodes a double‐stranded RNA‐binding protein, regulates leaf morphology and thermotolerance in rice through alteration of microRNA biogenesis. The srl10 mutant had a semi‐rolled leaf phenotype and elevated sensitivity to high temperature. SRL10 directly interacted with catalase isozyme B (CATB), and the two proteins mutually increased one other's stability to enhance hydrogen peroxide (H2O2) scavenging, thereby contributing to thermotolerance. The natural Hap3 (AGC) type of SRL10 allele was found to be present in the majority of aus rice accessions, and was identified as a thermotolerant allele under high temperature stress in both the field and the growth chamber. Moreover, the seed‐setting rate was 3.19 times higher and grain yield per plant was 1.68 times higher in near‐isogenic line (NIL) carrying Hap3 allele compared to plants carrying Hap1 allele under heat stress. Collectively, these results reveal a new locus of interest and define a novel SRL10–CATB based regulatory mechanism for developing cultivars with high temperature tolerance and stable yield. Furthermore, our findings provide a theoretical basis for simultaneous breeding for plant architecture and stress resistance. [ABSTRACT FROM AUTHOR]

Grain number per panicle (GNPP), determined mainly by panicle branching, is vital for rice yield. The dissection of the genetic basis underlying GNPP could help to improve rice yield. However, genetic resources, including quantitative trait loci (QTL) or genes for breeders to enhance rice GNPP, are still limited. Here, we conducted the genome-wide association study (GWAS) on the GNPP, primary branch number (PBN), and secondary branch number (SBN) of 468 rice accessions. We detected a total of 18 QTLs, including six for GNPP, six for PBN, and six for SBN, in the whole panel and the indica and japonica subpanels of 468 accessions. More importantly, qPSG1 was a common QTL for GNPP, PBN, and SBN and was demonstrated by chromosome segment substitution lines (CSSLs). Considering gene annotation, expression, and haplotype analysis, seven novel and strong GNPP-related candidate genes were mined from qPSG1. Our results provide clues to elucidate the molecular regulatory network of GNPP. The identified QTLs and candidate genes will contribute to the improvement of GNPP and rice yield via molecular marker-assisted selection (MAS) breeding and genetic engineering techniques. [ABSTRACT FROM AUTHOR]

Leaf morphology is one of the important traits related to ideal plant architecture and is an important factor determining rice stress resistance, which directly affects yield. Wax layers form a barrier to protect plants from different environmental stresses. However, the regulatory effect of wax synthesis genes on leaf morphology and salt tolerance is not well-understood. In this study, we identified a rice mutant, leaf tip rumpled 1 (ltr1), in a mutant library of the classic japonica variety Nipponbare. Phenotypic investigation of NPB and ltr1 suggested that ltr1 showed rumpled leaf with uneven distribution of bulliform cells and sclerenchyma cells, and disordered vascular bundles. A decrease in seed-setting rate in ltr1 led to decreased per-plant grain yield. Moreover, ltr1 was sensitive to salt stress, and LTR1 was strongly induced by salt stress. Map-based cloning of LTR1 showed that there was a 2-bp deletion in the eighth exon of LOC_Os02g40784 in ltr1, resulting in a frameshift mutation and early termination of transcription. Subsequently, the candidate gene was confirmed using complementation, overexpression, and knockout analysis of LOC_Os02g40784. Functional analysis of LTR1 showed that it was a wax synthesis gene and constitutively expressed in entire tissues with higher relative expression level in leaves and panicles. Moreover, overexpression of LTR1 enhanced yield in rice and LTR1 positively regulates salt stress by affecting water and ion homeostasis. These results lay a theoretical foundation for exploring the molecular mechanism of leaf morphogenesis and stress response, providing a new potential strategy for stress-tolerance breeding. [ABSTRACT FROM AUTHOR]

SUMMARY: Panicle development is an important determinant of the grain number in rice. A thorough characterization of the molecular mechanism underlying panicle development will lead to improved breeding of high‐yielding rice varieties. Frizzy Panicle (FZP), a critical gene for panicle development, is regulated by OsBZR1 and OsARFs at the transcriptional stage. However, the translational modulation of FZP has not been reported. We reveal that the CU‐rich elements (CUREs) in the 3′ UTR of the FZP mRNA are crucial for efficient FZP translation. The knockout of CUREs in the FZP 3′ UTR or the over‐expression of the FZP 3′ UTR fragment containing CUREs resulted in an increase in FZP mRNA translation efficiency. Moreover, the number of secondary branches (NSB) and the grain number per panicle (GNP) decreased in the transformed rice plants. The CUREs in the 3′ UTR of FZP mRNA were verified as the targets of the polypyrimidine tract‐binding proteins OsPTB1 and OsPTB2 in rice. Both OsPTB1 and OsPTB2 were highly expressed in young panicles. The knockout of OsPTB1/2 resulted in an increase in the FZP translational efficiency and a decrease in the NSB and GNP. Furthermore, the over‐expression of OsPTB1/2 decreased the translation of the reporter gene fused to FZP 3′ UTR in vivo and in vitro. These results suggest that OsPTB1/2 can mediate FZP translational repression by interacting with CUREs in the 3′ UTR of FZP mRNA, leading to changes in the NSB and GNP. Accordingly, in addition to transcriptional regulation, FZP expression is also fine‐tuned at the translational stage during rice panicle development. Significance Statement: Panicle architecture plays an important role in grain number determination in rice. Here, we identified a conserved RNA element, CU‐rich element (CURE), at the 3′‐untranslated region of Frizzy Panicle (FZP) which is a well‐studied gene regulating panicle development in rice. The CUREs could recruit the binding proteins OsPTB1/2 to repress the translation of FZP itself, which regulates the differentiation of secondary branches in panicle development. [ABSTRACT FROM AUTHOR]

Leaf and panicle are important nutrient and yield organs in rice, respectively. Although several genes controlling lesion mimic leaf and panicle abortion have been identified, a few studies have reported the involvement of a single gene in the production of both the traits. In this study, we characterized a panicle abortion mutant, lesion mimic leaf and panicle apical abortion (lmpa), which exhibits lesions on the leaf and causes degeneration of apical spikelets. Molecular cloning revealed that LMPA encodes a proton pump ATPase protein that is localized in the plasma membrane and is highly expressed in leaves and panicles. The analysis of promoter activity showed that the insertion of a fragment in the promoter of lmpa caused a decrease in the transcription level. Cellular and histochemistry analysis indicated that the ROS accumulated and cell death occurred in lmpa. Moreover, physiological experiments revealed that lmpa was more sensitive to high temperatures and salt stress conditions. These results provide a better understanding of the role of LMPA in panicle development and lesion mimic formation by regulating ROS homeostasis. [ABSTRACT FROM AUTHOR]

Summary: High‐temperature stress inhibits normal cellular processes and results in abnormal growth and development in plants. However, the mechanisms by which rice (Oryza sativa) copes with high temperature are not yet fully understood.In this study, we identified a rice high temperature enhanced lesion spots 1 (hes1) mutant, which displayed larger and more dense necrotic spots under high temperature conditions. HES1 encoded a UDP‐N‐acetylglucosamine pyrophosphorylase, which had UGPase enzymatic activity. RNA sequencing analysis showed that photosystem‐related genes were differentially expressed in the hes1 mutant at different temperatures, indicating that HES1 plays essential roles in maintaining chloroplast function. HES1 expression was induced under high temperature conditions.Furthermore, loss‐of‐function of HES1 affected heat shock factor expression and its mutation exhibited greater vulnerability to high temperature. Several experiments revealed that higher accumulation of reactive oxygen species occurred in the hes1 mutant at high temperature. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and comet experiments indicated that the hes1 underwent more severe DNA damage at high temperature. The determination of chlorophyll content and chloroplast ultrastructure showed that more severe photosystem defects occurred in the hes1 mutant under high temperature conditions.This study reveals that HES1 plays a key role in adaptation to high‐temperature stress in rice. [ABSTRACT FROM AUTHOR]

Summary: The riboflavin derivatives flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD) are essential cofactors for enzymes in multiple cellular processes. Characterizing mutants with impaired riboflavin metabolism can help clarify the role of riboflavin in plant development. Here, we characterized a rice (Oryza sativa) white and lesion‐mimic (wll1) mutant, which displays a lesion‐mimic phenotype with white leaves, chlorophyll loss, chloroplast defects, excess reactive oxygen species (ROS) accumulation, decreased photosystem protein levels, changes in expression of chloroplast development and photosynthesis genes, and cell death. Map‐based cloning and complementation test revealed that WLL1 encodes lumazine synthase, which participates in riboflavin biosynthesis. Indeed, the wll1 mutant showed riboflavin deficiency, and application of FAD rescued the wll1 phenotype. In addition, transcriptome analysis showed that cytokinin metabolism was significantly affected in wll1 mutant, which had increased cytokinin and δ‐aminolevulinic acid contents. Furthermore, WLL1 and riboflavin synthase (RS) formed a complex, and the rs mutant had a similar phenotype to the wll1 mutant. Taken together, our findings revealed that WLL1 and RS play pivotal roles in riboflavin biosynthesis, which is necessary for ROS balance and chloroplast development in rice. Significance Statement: WLL1 and RS play pivotal roles in riboflavin biosynthesis, which is necessary for ROS balance and chloroplast development in rice. [ABSTRACT FROM AUTHOR]

The genetic improvement of nitrogen use efficiency (NUE) of crops is vital for grain productivity and sustainable agriculture. However, the regulatory mechanism of NUE remains largely elusive. Here, we report that the rice Grain number, plant height, and heading date7 (Ghd7) gene genetically acts upstream of ABC1 REPRESSOR1 (ARE1), a negative regulator of NUE, to positively regulate nitrogen utilization. As a transcriptional repressor, Ghd7 directly binds to two Evening Element-like motifs in the promoter and intron 1 of ARE1 , likely in a cooperative manner, to repress its expression. Ghd7 and ARE1 display diurnal expression patterns in an inverse oscillation manner, mirroring a regulatory scheme based on these two loci. Analysis of a panel of 2656 rice varieties suggests that the elite alleles of Ghd7 and ARE1 have undergone diversifying selection during breeding. Moreover, the allelic distribution of Ghd7 and ARE1 is associated with the soil nitrogen deposition rate in East Asia and South Asia. Remarkably, the combination of the Ghd7 and ARE1 elite alleles substantially improves NUE and yield performance under nitrogen-limiting conditions. Collectively, these results define a Ghd7 – ARE1 -based regulatory mechanism of nitrogen utilization, providing useful targets for genetic improvement of rice NUE. This study reveals that the transcription factor Ghd7 directly represses the expression of ARE1 to positively regulate nitrogen use efficiency and grain yield in rice. While the allelic distribution of the Ghd7 – ARE1 regulatory module, which has undergone diversifying selection, is associated with the soil nitrogen deposition rate, the combination of their elite alleles significantly boosts rice productivity under low-nitrogen conditions. [ABSTRACT FROM AUTHOR]

Senescence in plants is induced by endogenous physiological changes and exogenous stresses. In this study, we isolated two alleles of a novel rice (Oryza sativa) mutant, yellow and premature dwarf 1 (ypd1). The ypd1 mutants exhibited a yellow and dwarf phenotype from germination, and premature senescence starting at tillering. Moreover, the ypd1 mutants were sensitive to high light, which accelerated cell death and senescence. Consistent with their yellow phenotype, the ypd1 mutants had abnormal chloroplasts and lower levels of photosynthetic pigments. TUNEL assays together with histochemical staining demonstrated that ypd1 mutants showed cell death and that they accumulated reactive oxygen species. The ypd1 mutants also showed increased expression of genes associated with senescence. Map-based cloning revealed a G→A substitution in exon 6 (ypd1-1) and exon 13 (ypd1-2) of LOC_Os06g13050 that affected splicing and caused premature termination of the encoded protein. YPD1 was found to be preferentially expressed in the leaf and it encodes a LRR-like1 protein. Complementation, overexpression, and targeted deletion confirmed that the mutations in YPD1 caused the ypd1 phenotype. YPD1 was localized on the chloroplast membrane. Our results thus demonstrate that the novel rice LRR-like1 protein YPD1 affects chloroplast development and leaf senescence. [ABSTRACT FROM AUTHOR]

Summary: Lesion‐mimic mutants (LMMs) provide a valuable tool to reveal the molecular mechanisms determining programmed cell death (PCD) in plants. Despite intensive research, the mechanisms behind PCD and the formation of lesions in various LMMs still remain to be elucidated. Here, we identified a rice (Oryza sativa) LMM, early lesion leaf 1 (ell1), cloned the causal gene by map‐based cloning, and verified this by complementation. ELL1 encodes a cytochrome P450 monooxygenase, and the ELL1 protein was located in the endoplasmic reticulum. The ell1 mutant exhibited decreased chlorophyll contents, serious chloroplast degradation, upregulated expression of chloroplast degradation‐related genes, and attenuated photosynthetic protein activity, indicating that ELL1 is involved in chloroplast development. RNA sequencing analysis showed that genes related to oxygen binding were differentially expressed in ell1 and wild‐type plants; histochemistry and paraffin sectioning results indicated that hydrogen peroxide (H2O2) and callose accumulated in the ell1 leaves, and the cell structure around the lesions was severely damaged, which indicated that reactive oxygen species (ROS) accumulated and cell death occurred in the mutant. TUNEL staining and comet experiments revealed that severe DNA degradation and abnormal PCD occurred in the ell1 mutants, which implied that excessive ROS accumulation may induce DNA damage and ROS‐mediated cell death in the mutant. Additionally, lesion initiation in the ell1 mutant was light dependent and temperature sensitive. Our findings revealed that ELL1 affects chloroplast development or function, and that loss of ELL1 function induces ROS accumulation and lesion formation in rice. Significance Statement: ELL1 affects chloroplast development or function, and loss of ELL1 function induces ROS accumulation and lesion formation in rice. [ABSTRACT FROM AUTHOR]

Summary: The biosynthesis and modification of cell wall composition and structure are controlled by hundreds of enzymes and have a direct consequence on plant growth and development. However, the majority of these enzymes has not been functionally characterised.Rice mutants with leaf‐rolling phenotypes were screened in a field. Phenotypic analysis under controlled conditions was performed for the selected mutant and the relevant gene was identified by map‐based cloning. Cell wall composition was analysed by glycome profiling assay.We identified a photo‐sensitive leaf rolling 1 (psl1) mutant with 'napping' (midday depression of photosynthesis) phenotype and reduced growth. The PSL1 gene encodes a cell wall‐localised polygalacturonase (PG), a pectin‐degrading enzyme. psl1 with a 260‐bp deletion in its gene displayed leaf rolling in response to high light intensity and/or low humidity. Biochemical assays revealed PG activity of recombinant PSL1 protein. Significant modifications to cell wall composition in the psl1 mutant compared with the wild‐type plants were identified. Such modifications enhanced drought tolerance of the mutant plants by reducing water loss under osmotic stress and drought conditions.Taken together, PSL1 functions as a PG that modifies cell wall biosynthesis, plant development and drought tolerance in rice. [ABSTRACT FROM AUTHOR]

The juvenile‐to‐adult transition in plants involves changes in vegetative growth and plant architecture; the timing of this transition has important implications for agriculture. The microRNA miR156 regulates this transition and shoot maturation in plants. In Arabidopsis thaliana, deposition of histone H3 trimethylation on lysine 27 (H3K27me3, a repressive mark) at the MIR156A/C loci is regulated by Polycomb Repressive Complex 1 (PRC1) or PRC2, depending on the developmental stage. The levels of miR156 progressively decline during shoot maturation. The amount of H3K27me3 at MIR156A/C loci affects miR156 levels; however, whether this epigenetic regulation is conserved remains unclear. Here, we found that in rice (Oryza sativa), the putative PRC1 subunit LIKE HETEROCHROMATIN PROTEIN 1 (OsLHP1), with the miR156–SQUAMOSA PROMOTER BINDING PROTEIN‐LIKE (SPL) module, affects developmental phase transitions. Loss of OsLHP1 function results in ectopic expression of MIR156B/C/I/E, phenocopy of miR156 overexpression, and reduced H3k27me3 levels at MIR156B/C/I/E. This indicates that OsLHP1 has functionally diverged from Arabidopsis LHP1. Genetic and transcriptome analyses of wild‐type, miR156b/c‐overexpression, and Oslhp1‐2 mutant plants suggest that OsLHP1 acts upstream of miR156 and SPL during the juvenile‐to‐adult transition. Therefore, modifying the OsLHP1–miR156–SPL pathway may enable alteration of the vegetative period and plant architecture. [ABSTRACT FROM AUTHOR]

SUMMARY: Ferredoxins (Fds) play a crucial role in photosynthesis by regulating the distribution of electrons to downstream enzymes. Multiple Fd genes have been annotated in the Oryza sativa L. (rice) genome; however, their specific functions are not well understood. Here, we report the functional characterization of rice Fd1. Sequence alignment, phylogenetic analysis of seven rice Fd proteins and quantitative reverse transcription polymerase chain reaction (qRT‐PCR) analysis showed that rice Fd1 is a primary leaf‐type Fd. Electron transfer assays involving NADP+ and cytochrome c indicated that Fd1 can donate electrons from photosystem I (PSI) to ferredoxin‐NADP+ reductase. Loss‐of‐function fd1 mutants showed chlorosis and seedling lethality at the three‐leaf stage. The deficiency of Fd1 impaired photosynthetic electron transport, which affected carbon assimilation. Exogenous glucose treatment partially restored the mutant phenotype, suggesting that Fd1 plays an important role in photosynthetic electron transport in rice. In addition, the transcript levels of Fd‐dependent genes were affected in fd1 mutants, and the trend was similar to that observed in fdc2 plants. Together, these results suggest that OsFd1 is the primary Fd in photosynthetic electron transport and carbon assimilation in rice. Significance Statement: Ferredoxins (Fds) play a crucial role in photosynthesis by regulating the distribution of electrons to downstream enzymes; however, the specific functions of multiple Fd genes annotated in the rice genome are not well understood. This study revealed that disruption of rice Fd1, encoding a primary leaf‐type ferredoxin that functions in photosynthetic electron transport, leads to chlorosis and seedling lethality, and that the expression of Fd1 and FdC2 are coordinated in rice. [ABSTRACT FROM AUTHOR]

Summary: Understanding the genetic basis of natural variation in grain size among diverse rice varieties can help breeders develop high‐yielding rice cultivars.Here, we report the discovery of qTGW2, a new semidominant quantitative trait locus for grain width and weight. The corresponding gene, TGW2, encodes CELL NUMBER REGULATOR 1 (OsCNR1) localized to the plasma membrane.A single nucleotide polymorphism (SNP) variation 1818 bp upstream of TGW2 is responsible for its different expression, leading to alteration in grain width and weight by influencing cell proliferation and expansion in glumes. TGW2 interacts with KRP1, a regulator of cell cycle in plants, to negatively regulate grain width and weight. Genetic diversity analysis of TGW2 in 141 rice accessions revealed it as a breeding target in a selective sweep region.Our findings provide new insights into the genetic mechanism underlying grain morphology and grain weight, and uncover a promising gene for improving rice yield. [ABSTRACT FROM AUTHOR]

Background: Copy number variations (CNVs) are an important type of structural variations in the genome that usually affect gene expression levels by gene dosage effect. Understanding CNVs as part of genome evolution may provide insights into the genetic basis of important agricultural traits and contribute to the crop breeding in the future. While available methods to detect CNVs utilizing next-generation sequencing technology have helped shed light on prevalence and effects of CNVs, the complexity of crop genomes poses a major challenge and requires development of additional tools. Results: Here, we generated genomic and transcriptomic data of 93 rice (Oryza sativa L.) accessions and developed a comprehensive pipeline to call CNVs in this large-scale dataset. We analyzed the correlation between CNVs and gene expression levels and found that approximately 13% of the identified genes showed a significant correlation between their expression levels and copy numbers. Further analysis showed that about 36% of duplicate pairs were involved in pseudogenetic events while only 5% of them showed functional differentiation. Moreover, the offspring copy mainly contributed to the expression levels and seemed more likely to become a pseudogene, whereas the parent copy tended to maintain the function of ancestral gene. Conclusion: We provide a high-accuracy CNV dataset that will contribute to functional genomics studies and molecular breeding in rice. We also showed that gene dosage effect of CNVs in rice is not exponential or linear. Our work demonstrates that the evolution of duplicated genes is asymmetric in both expression levels and gene fates, shedding a new insight into the evolution of duplicated genes. [ABSTRACT FROM AUTHOR]

Plant architecture is a complex agronomic trait and a major factor of crop yield, which is affected by several important hormones. Strigolactones (SLs) are identified as a new class hormoneinhibiting branching in many plant species and have been shown to be involved in various developmental processes. Genetical and chemical modulation of the SL pathway is recognized as a promising approach to modify plant architecture. However, whether and how the genes involved in the SL pathway could be utilized in breeding still remain elusive. Here, we demonstrate that a partial loss-of-function allele of the SL biosynthesis gene, HIGH TILLERING AND DWARF 1 / DWARF17 (HTD1 / D17), which encodes CAROTENOID CLEAVAGE DIOXYGENASE 7 (CCD7), increases tiller number and improves grain yield in rice. We found that the HTD1 gene had been widely utilized and co-selected with Semidwarf 1 (SD1), both contributing to the improvement of plant architecture in modern rice varieties since the Green Revolution in the 1960s. Understanding how phytohormone pathway genes regulate plant architecture and how they have been utilized and selected in breeding will lay the foundation for developing the rational approaches toward improving crop yield. A partial loss-of-function allele of strigolactone biosynthesis gene HTD1 , which increases tiller number and enhances grain yield, was identified in a widely cultivated high-yield rice variety, Huazhan. This beneficial allele of HTD1 (HTD1 HZ ) was found to be co-selected with the beneficial allele of SD1 (SD1 DGWG ) by breeders during the Green Revolution and together passed into modern elite varieties. [ABSTRACT FROM AUTHOR]

Summary: Heading date 1 (Hd1) is an important gene for the regulation of flowering in rice, but its variation in major cultivated rice varieties, and the effect of this variation on yield and quality, remains unknown. In this study, we selected 123 major rice varieties cultivated in China from 1936 to 2009 to analyse the relationship between the Hd1 alleles and yield‐related traits. Among these varieties, 19 haplotypes were detected in Hd1, including two major haplotypes (H8 and H13) in the japonica group and three major haplotypes (H14, H15 and H16) in the indica group. Analysis of allele frequencies showed that the secondary branch number was the major aimed for Chinese indica breeding. In the five major haplotypes, SNP316(C‐T) was the only difference between the two major japonica haplotypes, and SNP495(C‐G) and SNP614(G‐A) are the major SNPs in the three indica haplotypes. Association analysis showed that H16 is the most preponderant allele in modern cultivated Chinese indica varieties. Backcrossing this allele into the japonica variety Chunjiang06 improved yield without decreasing grain quality. Therefore, our analysis offers a new strategy for utilizing these preponderant alleles to improve yield and quality of japonica varieties for cultivation in the southern areas of China. [ABSTRACT FROM AUTHOR]

Keywords: rice; biomass; grain number; nitrogen utilization efficiency; grain yield EN rice biomass grain number nitrogen utilization efficiency grain yield 1681 1683 3 09/13/21 20210901 NES 210901 Since the "first green revolution" in the 1960s, rice grain yield has risen sharply. The SEEDLING BIOMASS 1 allele from indica rice enhances yield performance under low-nitrogen environments Meanwhile, given its another characteristic of enhancing nitrogen utilization, I SBM1 i would improve yield performance at low-nitrogen supply, especially if introgressing the HapB of I SBM1 i to I japonica i rice that lacks this favourable haplotype. [Extracted from the article]

Cadmium (Cd) is one of the most toxic heavy metals, and its accumulation in plants will seriously affect growth and yield. In this study, Cd-sensitive line D69 and Cd-tolerant line D28 were selected, which the Cd content of D28 was higher than D69 in both above and underground parts after Cd treatment. Using a combination of two-dimensional gel electrophoresis (2-DE) and MALDI-TOF-TOF MS/MS, the differential expression changes of phosphorylated proteins between D69 and D28 in leaves were classified and analyzed after Cd treatment. A total of 53 differentially expressed phosphoproteins were identified, which mainly involved in metabolism, signal transduction, gene expression regulation, material transport, and membrane fusion. The phosphorylated proteins of Cd-tolerant and Cd-sensitive lines were all analyzed, and found that some proteins associated with carbon metabolism, proteolytic enzymes, F-box containing transcription factors, RNA helicases, DNA replication/transcription/repair enzymes and ankyrins were detected in Cd-tolerant line D28, which might alleviate the abiotic stress caused by Cd treatment. These results will clarify the phosphorylated pathways in response and resistance to Cd stress in rice. • Protein phosphorylation plays an important role in responses to Cd stress. • General response mechanisms to resist Cd toxicity exist in Cd-sensitive and Cd-tolerant rice lines. • Resistant mechanisms in Cd-tolerant line might alleviate the abiotic stress caused by Cd treatment. [ABSTRACT FROM AUTHOR]

Summary: ATP‐citrate lyases (ACL) play critical roles in tumour cell propagation, foetal development and growth, and histone acetylation in human and animals. Here, we report a novel function of ACL in cell death‐mediated pathogen defence responses in rice. Using ethyl methanesulphonate (EMS) mutagenesis and map‐based cloning, we identified an Oryza sativa ACL‐A2 mutant allele, termed spotted leaf 30‐1 (spl30‐1), in which an A‐to‐T transversion converts an Asn at position 343 to a Tyr (N343Y), causing a recessive mutation that led to a lesion mimic phenotype. Compared to wild‐type plants, spl30‐1 significantly reduces ACL enzymatic activity, accumulates high reactive oxygen species and increases degradation rate of nuclear deoxyribonucleic acids. CRISPR/Cas9‐mediated insertion/deletion mutation analysis and complementation assay confirmed that the phenotype of spl30‐1 resulted from the defective function of OsACL‐A2 protein. We further biochemically identified that the N343Y mutation caused a significant degradation of SPL30N343Y in a ubiquitin‐26S proteasome system (UPS)‐dependent manner without alteration in transcripts of OsACL‐A2 in spl30‐1. Transcriptome analysis identified a number of up‐regulated genes associated with pathogen defence responses in recessive mutants of OsACL‐A2, implying its role in innate immunity. Suppressor mutant screen suggested that OsSL, which encodes a P450 monooxygenase protein, acted as a downstream key regulator in spl30‐1‐mediated pathogen defence responses. Taken together, our study discovered a novel role of OsACL‐A2 in negatively regulating innate immune responses in rice. [ABSTRACT FROM AUTHOR]

Summary: Water deficit is a major environmental threat affecting crop yields worldwide. In this study, a drought stress‐sensitive mutant drought sensitive 8 (ds8) was identified in rice (Oryza sativa L.). The DS8 gene was cloned using a map‐based approach. Further analysis revealed that DS8 encoded a Nck‐associated protein 1 (NAP1)‐like protein, a component of the SCAR/WAVE complex, which played a vital role in actin filament nucleation activity. The mutant exhibited changes in leaf cuticle development. Functional analysis revealed that the mutation of DS8 increased stomatal density and impaired stomatal closure activity. The distorted actin filaments in the mutant led to a defect in abscisic acid (ABA)‐mediated stomatal closure and increased ABA accumulation. All these resulted in excessive water loss in ds8 leaves. Notably, antisense transgenic lines also exhibited increased drought sensitivity, along with impaired stomatal closure and elevated ABA levels. These findings suggest that DS8 affects drought sensitivity by influencing actin filament activity. Significance Statement: DS8 encodes a NAP1‐like protein that affects drought sensitivity by its involvement in leaf epidermal development and stomatal closure in rice, revealing a molecular mechanism concerning the SCAR/WAVE‐Arp2/3 complex in the drought stress response. [ABSTRACT FROM AUTHOR]

Pentatricopeptide repeat (PPR) proteins regulate organellar gene expression in plants, through their involvement in organellar RNA metabolism. In rice (Oryza sativa), 477 genes are predicted to encode PPR proteins; however, the majority of their functions remain unknown. In this study, we identified and characterized a rice mutant, pale-green leaf12 (pgl12); at the seedling stage, pgl12 mutants had yellow-green leaves, which gradually turned pale green as the plants grew. The pgl12 mutant had significantly reduced Chl contents and increased sensitivity to changes in temperature. A genetic analysis revealed that the pgl12 mutation is recessive and located within a single nuclear gene. Map-based cloning of PGL12, including a transgenic complementation test, confirmed the presence of a base substitution (C to T), generating a stop codon, within LOC_Os12g10184 in the pgl12 mutant. LOC_Os12g10184 encodes a novel PLS-type PPR protein containing 17 PPR motifs and targeted to the chloroplasts. A quantitative real-time PCR analysis showed that PGL12 was expressed in various tissues, especially the leaves. We also showed that the transcript levels of several nuclear- and plastid-encoded genes associated with chloroplast development and photosynthesis were significantly altered in pgl12 mutants. The mutant exhibited defects in the 16S rRNA processing and splicing of the plastid transcript ndhA. Our results indicate that PGL12 is a new PLS-type PPR protein required for proper chloroplast development and 16S rRNA processing in rice. [ABSTRACT FROM AUTHOR]

Abstract Chloroplast transit peptides (CTPs) can be used to transport non-chloroplastic proteins into the chloroplasts. Here, we studied the CTPs of three rice (Oryza sativa L.) chloroplast-localized proteins and found that their CTPs could be used to transport non-chloroplast-localized proteins into the chloroplasts. Fusion proteins lacking the CTP remained located in the cytoplasm. Furthermore, we constructed green fluorescent protein fusion vectors with the three CTPs and three non-chloroplast-localized proteins, Ghd10, MULTI-FLORET SPIKELET1 (MFS1), and SHORTENED UPPERMOST INTERNODE 1 (SUI1). After transforming these constructs into rice protoplasts, the fusion proteins all localized in the chloroplasts. Collectively, our results showed that these CTPs can transport non-chloroplast-localized proteins into the chloroplasts, and more importantly, these CTPs can be applied to engineer chloroplast metabolism. [ABSTRACT FROM AUTHOR]

Ferredoxin (Fd) protein as unique electron acceptor, involved in a variety of fundamental metabolic and signaling processes, which is indispensable for plant growth. The molecular mechanisms of Fd such as regulation of electron partitioning, impact of photosynthetic rate and involvement in the carbon fixing remain elusive in rice. Here we reported a heading date delay and yellowish leaf 1 (hdy1) mutant derived from Japonica rice cultivar “Nipponbare” subjected to EMS treatment. In the paddy field, the hdy1 mutant appeared at a significantly late heading date and had yellow-green leaves during the whole growth stage. Further investigation indicated that the abnormal phenotype of hdy1 was connected with depressed pigment content and photosynthetic rate. Genetic analysis results showed that the hdy1 mutant phenotype was caused by a single recessive nuclear gene mutation. Map-based cloning revealed that OsHDY1 is located on chromosome 3 and encodes an ortholog of the AtFdC2 gene. Complementation and overexpression, transgenic plants exhibited the mutant phenotype including head date, leaf color and the transcription levels of the FdC2 were completely rescued by transformation with OsHDY1. Real-time PCR revealed that the expression product of OsHDY1 was detected in almost all of the organs except root, whereas highest expression levels were observed in seeding new leaves. The lower expression levels of HDY1 and content of iron were detected in hdy1 than WT’s. The FdC2::GFP was detected in the chloroplasts of rice. Real-time PCR results showed that the expression of many photosynthetic electron transfer related genes in hdy1 were higher than WT. Our results suggest that OsFdC2 plays an important role in photosynthetic rate and development of heading date by regulating electron transfer and chlorophyll content in rice. [ABSTRACT FROM AUTHOR]

Grain size determines grain weight and affects grain quality. Several major quantitative trait loci (QTLs) regulating grain size have been cloned; however, our understanding of the underlying mechanism that regulates the size of rice grains remains fragmentary. Here, we report the cloning and characterization of a dominant QTL, GRAIN SIZE ON CHROMOSOME 2 ( GS2 ), which encodes Growth-Regulating Factor 4 (OsGRF4), a transcriptional regulator. GS2 localizes to the nucleus and may act as a transcription activator. A rare mutation of GS2 affecting the binding site of a microRNA, OsmiR396c, causes elevated expression of GS2 / OsGRF4 . The increase in GS2 expression leads to larger cells and increased numbers of cells, which thus enhances grain weight and yield. The introduction of this rare allele of GS2 / OsGRF4 into rice cultivars could significantly enhance grain weight and increase grain yield, with possible applications in breeding high-yield rice varieties. [ABSTRACT FROM AUTHOR]

The aerial parts of higher plants are generated from the shoot apical meristem (SAM). In this study, we isolated a small rice (Oryza sativa L.) mutant that showed premature termination of shoot development and was named mini rice 1 (mini1). The mutant was first isolated from a japonica cultivar Zhonghua11 (ZH11) subjected to ethyl methanesulfonate (EMS) treatment. With bulked segregant analysis (BSA) and mapbased cloning method, Mini1 gene was finally fine-mapped to an interval of 48.6 kb on chromosome 9. Sequence analyses revealed a single base substitution from G to A was found in the region, which resulted in an amino acid change from Gly to Asp. The candidate gene Os09g0363900 was predicted to encode a putative adhesion of calyx edges protein ACE (putative HOTHEAD precursor) and genetic complementation experiment confirmed the identity of Mini1. Os09g0363900 contains glucose-methanol-choline (GMC) oxidoreductase and NAD(P)- binding Rossmann-like domain, and exhibits high similarity to Arabidopsis HOTHEAD (HTH). Expression analysis indicated Mini1 was highly expressed in young shoots but lowly in roots and the expression level of most genes involved in auxin biosynthesis and signal transduction were reduced in mutant. We conclude that Mini1 plays an important role in maintaining SAM activity and promoting shoot development in rice. [ABSTRACT FROM AUTHOR]

Abstract: Molecular design breeding is one of straightforward approaches to break yield barriers in rice. In this study, GW6 gene for grain length and width from Baodali was transferred into an indica recurrent parent 9311 and a japonica variety Zhonghua 11 (ZH11) using marker-assisted backcross (MAB). One and three introgression lines were selected for phenotypic analysis from 9311 and ZH11 genetic backgrounds, respectively. SSL-1, an improved 9311 near isogenic line with GW6 performed 11%, 19% and 6.7% higher of grain length, 1000-grain weight and single plant yield, respectively, as compared with 9311. All the three improved ZH11-GW6 lines, R1, R2 and R3, had more than 30% increase in grain weight and about 7% higher in grain yield. Seed plumpness of R1, R2 and R3 was improved synchronously because the three ZH11-GW6 lines contained GIF1 (Grain Incomplete Filling 1), a dominant grain filling gene. Thus, GW6 has high potential in increasing the yield of inbred lines through MAB, making it an important genetic resource in super hybrid rice breeding. This study provides insights in the utilization of GW6 for large grain and high yield rice breeding via molecular design breeding. [Copyright &y& Elsevier]

The increasing living standard of customers has resulted in a demand for the improvement of the cooking, eating, and appearance quality of rice grains, which have become a priority for rice breeders and geneticists. In this study, a double haploid population derived from the cross between japonica CJ06 and indica TN1 was used to analyze the quantitative trait loci (QTLs) for amylose content, gel consistency (GC), gelatinization temperature (GT), protein content, and grain hardness under two different conditions. A total of 18 QTLs were detected on chromosomes 1, 2, 3, 6, 8, 10, and 12, with the additive heritability ranging from 10.4 to 66.3 %. Eight of them were identified both in Hangzhou and Hainan. Moreover, five-locus epistatic interactions were identified except for GC, and two QTLs for GT exhibited environmental interaction effects. The results facilitate further understanding of the genetic basis for eating and cooking quality, nutritive quality, and milling quality. [ABSTRACT FROM AUTHOR]

Proper flowering time is essential for plant reproduction. Winter annual Arabidopsis thaliana needs vernalization before flowering, during which AtVILs (VIN3 and VRN5, components of PRC2 complex) mediate the H3K27 tri-methylation at the FLC locus (a floral repressor) to repress the FLC expression and hence to induce flowering. However, how VILs (VIL, VERNALIZATION INSENSITIVE 3-LIKE) function in rice is unknown. Here we demonstrated that rice LC2 (OsVIL3) and OsVIL2 (two OsVILs, possible components of PRC2 complex) promote rice flowering. Our results showed that expressions of LC2 and OsVIL2 are induced by SD (short-day) conditions and both lc2 mutant and OsVIL2–RNAi lines display delayed heading date, consistent with the reduced expression levels of Hd1 and Hd3a. Interestingly, LC2 binds to the promoter region of a floral repressor OsLF and represses the OsLF expression via H3K27 tri-methylation modification. In addition, OsLF directly regulates the Hd1 expression through binding to Hd1 promoter. These results first demonstrated that the putative PRC2 in rice is involved in photoperiod flowering regulation, which is different from that of Arabidopsis, and revealed that LC2 binds the promoter region of target gene, presenting a possible mechanism of the recruitment process of PRC2 complex to its target genes. The studies provide informative clues on the epigenetic control of rice flowering. [ABSTRACT FROM PUBLISHER]

Abstract: The extraction of DNA is often the most time consuming and laborious step in high-throughput molecular genetic analysis and marker assisted selection (MAS) programs. A simple method for preparation of rice genomic DNA was developed. A small amount (1-50 mg) of leaf tissue of rice seedling, 500 μL of extraction buffer, and one steel bead were put into a 2-mL microcentrifuge tube. After vigorously mashing for 2 min, 5 μL of supernatant was directly applied to PCR amplification. Otherwise, the supernatant was precipitated with two times volume of ethanol to obtain high quality genomic DNA. This method is simple, rapid, low cost, and reliable for PCR analysis. One person can manipulate as many as 96 samples for PCR in 10 min. It is especially suitable for genotyping of large number of samples. [ABSTRACT FROM AUTHOR]