Sterile Rice Research Articles

Screening of secondary metabolites produced by a mangroove-derived Nigrospora species an endophytic fungus isolated from Rhizophora racemosa for antioxidant and antimicrobial properties

Marine fungi have shown large chemo-diversity of unharnessed important secondary metabolites required for drug development. In this study, endophytic fungi Nigrospora species (5S3) was isolated from the fresh stem of Rhizophora racemosa from a mangrove forest in Lagos Nigeria and axenic colonies were fermented on sterile rice medium for 21 days. The antimicrobial and antioxidant activities of the extract were evaluated. Chemical analyses such as High Performance Liquid Chromatography Diode Array Detector (HPLC-DAD) and Nuclear magnetic Resonance Imaging (NMR) analyses of selected Vacuum Liquid Chromatography (VLC) fractions revealed the presence of ten known compounds with established biological activities. Tested fractions of the extract exhibited antimicrobial activities against at least one Gram positive and Gram negative bacteria with MIC values that ranged between 0.13 to 1 mg/mL. FE2 and FE3 fractions of the extract demonstrated poor antioxidant activity of less than 10%. Septicine, Aureonitol, Papuamine, Di-iso-octylphtalat, 1-(2,4 Dihydroxy-3,5-Dimethylphenyl)-Ethanone, Cladosporin, Tetrabenzofuran, Dihydrophthalate, 9-Octadecaenoic acid and Eicosane were the compounds detected in the extracts. Our findings reveal that Nigrospora spp an endophytic fungi possesses unique chemo diversity of bioactive secondary metabolites that could be utilized for development of new drugs.

Genetic Variability of Floral and Agronomic Characteristics that Influence Outcrossing Rate Percentage of Cytoplasmic Male Sterile Rice

Developing high-yielding hybrid rice varieties is much needed to ensure food security to meet the requirement of the increasing population across the world. Cytoplasmic male genetic sterility system is a valuable technology for the exploitation of heterosis and production of high-yielding hybrid rice with better grain quality. Large-scale commercial exploitation of hybrid vigor in rice largely depends on the availability of good CMS (Cytoplasmic Male Sterile) lines. A total of eighteen traits of five CMS lines of rice were evaluated to study the variability and correlation between floral and other yield-contributing traits with outcrossing rate using Randomized Complete Block Design with three replications. High significant variation was observed among the genotypes for all the studied traits. High GCV (Genotypic Coefficient of Variance) and PCV (phenotypic Coefficient of Variance) coupled with high heritability and high genetic advance in percent of mean were observed for number of panicles per plant, filled grain, flag leaf breadth, stigma length and breadth, filament length and outcrossing rate which indicated the broad genetic base, less environmental influence, and these traits are under the control of additive gene action. character association and path analysis indicated that number of grains per panicle, filled grains per panicle, stigma length, and breadth possessed both positive significant correlation and positive direct effects on outcrossing rate therefore, selection for the traits mentioned above will certainly bring in the improvement of the outcrossing percentage of the lines, which in turn enhances the seed quantity of the hybrid.

Base Editing of EUI1 Improves the Elongation of the Uppermost Internode in Two-Line Male Sterile Rice Lines

The use of male sterile lines (MSLs) of rice is essential for heterosis utilization. However, MSLs have a common defect in the elongation of the uppermost internode, eventually leading to incomplete panicle exsertion, blocking pollination, and reducing the hybrid rice seed yield. Previously, the elongated uppermost internode 1 (EUI1) was identified as an active gibberellin-deactivating enzyme that plays a key role in panicle exsertion from the flag leaf sheath in rice (Oryza sativa L.). We used an adenine base editor to edit EUI1 and obtained two types of homozygous transgenic plants (eui1-1 and eui1-2). The transcription and translation levels of EUI1 in the two mutants were significantly lower than in the wild type, as was the oxidation activity of EUI1 to active gibberellins (GAs), which also decreased. The contents of the plant hormones GA1, GA3, and GA4 in eui1-1 (1.64, 1.55, and 0.92 ng/g) and eui1-2 (0.85, 0.64, and 0.65 ng/g) panicles were significantly higher than the wild type (0.70, 0.57, and 0.42 ng/g). The uppermost internode lengths of the mutant were 26.5 and 23.6 cm, which were significantly longer than that of the wild type (18.0 cm), and the cell lengths of the mutant were 161.10 and 157.19 μm, which were longer than that of the wild type (89.28 μm). Our results indicate that the adenine base editing system could increase the content of endogenous bioactive GAs in young panicles by fine-tuning EUI1 activity, reduce the defect of panicle enclosure in MSLs and increase the yield of hybrid rice seed production.

Yuan Longping, a great world hunger fighter

Yuan Longping, a great world hunger fighter

Effect of sterile rice spikelets derived biochar amendment on nutrient leaching and availability in paddy soil under continuous standing water

ABSTRACT Undisturbed soil columns were used to study the effect of sterile rice spikelets-derived biochar (SRSB) on NH4 +-N, available P, and available K leaching and nutrients availability in a paddy soil under continuous standing water (CSW) condition. The experiment was conducted for two crop cycles and comprised of three treatments: recommended dose of sole chemical fertilizers (CF), CF+SRSB (2 t ha−1), and control. In both crop cycles, incorporation of SRSB reduced the leachate NH4 +-N and available K concentrations, while increased the available P concentration compared to sole CF. The SRSB amendment reduced the cumulative losses of NH4 +-N by 35–38% and available K by 23–25% than CF only. However, there was no significant variation in cumulative available P loss between SRSB and CF only. The reduced leaching loss of NH4 +-N and available K could be attributed to the better nutrient adsorption capacity of SRSB. After completion of two crop cycles, the soil pH, soil organic carbon, soil total N, available P, and available K increased with SRSB amended soils than sole CF. This study showed positive effects of SRSB on retaining nutrients and increasing their availability in the soil which could benefit efficient fertilizer management in paddy soil under CSW.

Environment-sensitive genic male sterility in rice and other plants.

Environment-sensitive genic male sterility is a type of male sterility that is affected by both genetic and environmental factors. Environment-sensitive genic male sterile lines are not only used in two-line hybrid breeding but are also good materials for studying plant-environment interactions. In this study we review the research progress on environment-sensitive genic male sterility in rice from the perspectives of epigenetic, transcriptional, posttranscriptional, posttranslationaland metabolic mechanisms as well as signal transduction processes. While significant progress has been made in the genetics, gene cloningand understanding of the molecular mechanisms of environment-sensitive genic male sterility in recent years, the relevant regulatory network is still poorly understood in rice. We therefore also review studies of environment-sensitive genic male sterility in Arabidopsis and other crops, hoping to promote research in this field in rice. Finally, we analyse the challenges posed by environment-sensitive genic male sterility and provide corresponding suggestions. This review will contribute towards an understanding the molecular genetics of environment-sensitive genic male sterility and its application in two-line hybrid breeding in rice and other species.

Rice anther tapetum: a vital reproductive cell layer for sporopollenin biosynthesis and pollen exine patterning.

The tapetum is the innermost layer of the four layers of the rice anther that provides protection and essential nutrients to pollen grain development and delivers precursors for pollen exine formation. The tapetum has a key role in the normal development of pollen grains and tapetal programmed cell death (PCD) that is linked with sporopollenin biosynthesis and transport. Recently, many genes have been identified that are involved in tapetum formation in rice and Arabidopsis. Genetic mutation in PCD-associated genes could affect normal tapetal PCD, which finally leads to aborted pollen grains and male sterility in rice. In this review, we discuss the most recent research on rice tapetum development, including genomic, transcriptomic and proteomic studies. Furthermore, tapetal PCD, sporopollenin biosynthesis, ROS activity for tapetum function and its role in male reproductive development are discussed in detail. This will improve our understanding of the role of the tapetum in male fertility using rice as a model system, and provide information that can be applied in rice hybridization and that of other major crops.

Disruption of the Expression of the Cinnamoyl–CoA Reductase (CCR) Gene OsCCR18 Causes Male Sterility in Rice (Oryza sativa L. japonica)

The biological process of anther development is very complex. It remains largely unclear how the cinnamoyl–CoA reductase (CCR) encoding genes function in the regulation of anther development in plants. Here, we establish that the CCR family gene OsCCR18 is essential for maintaining male fertility in rice. The OsCCR18 transcripts were greatly abundant in the panicles at the S4 and S5 developmental stages in rice. The subcellular localization of OsCCR18 proteins was in the nucleus of the rice. The knockout of the OsCCR18 gene resulted in a severely abnormal degradation of the tapetum as well as the abnormal development of granular Ubisch bodies, leading to the inability to form normal pollen in the mutants. Compared with the wild–type (WT) rice, the osccr18 mutants had no visible pollen grains and had entirely male sterility. Furthermore, several anther development–related genes, including OsPDA1, OsDTD, OsC6, OsACOS12, OsTDR, OsWDA1, OsDPW, OsCYP703A3, and OsNOP, were significantly lower expressed in the panicles at the stages from S5 to S8 in the osccr18 mutants than in the WT plants. Additionally, hundreds of genes involved in phenylpropanoid biosynthesis, fatty acid synthesis and metabolism exhibited distinct expression patterns between the WT and mutants, which may be crucial for controlling anther development in rice. These findings add a new regulatory role to CCR family gene–mediated male fertility in rice.

Loss of Function of the RRMF Domain in OsROS1a Causes Sterility in Rice (Oryza sativa L.).

For crop seed production, the development of anthers and male fertility are the main agronomic traits and key biological processes for flowering plants. Active DNA demethylation regulates many plant developmental processes and is ensured by 5-meC DNA glycosylase enzymes. To find out the role of OsROS1a, OsROS1a gene editing mutants were generated using the CRISPR/Cas9 system. The osros1a mutants had shrink spikelets, smaller anthers and pollen grains, and were not stained by iodine staining showing a significant reduction in total soluble sugar and starch contents as compared to wildtype (WT), which caused complete male sterility. Similarly, the expression of genes involved in pollen and anther development was decreased in osros1a mutants as compared to WT. Furthermore, bisulfite sequencing showed that the CG and CHG methylation of the OsPKS2 gene promoter was significantly increased in the osros1a mutant, which caused a reduced expression of OsPKS2 in osros1a mutants. DNA methylation of the TDR gene promoter was similar between WT and osros1a mutants, indicating that the DNA methylation effect by OsROS1a was gene specific. The expression of OsROS1a in the mutants was not changed, but it produced a frame-shift mutation to truncate the Pem-CXXC and RRMF domains. Combined with previous studies, our findings suggested that the RRMF domain in OsROS1a is the functional domain and loss of RRMF for OsROS1a causes sterility in rice.

Temperature-sensitive male sterility in rice determined by the roles of AGO1d in reproductive phasiRNA biogenesis and function.

Phased secondary siRNAs (phasiRNAs) are broadly present in the reproductive tissues of flowering plants, with spatial-temporal specificity. However, the ARGONAUTE (AGO) proteins associated with phasiRNAs and their miRNA triggers remain elusive. Here, through histological and high-throughput sequencing analyses, we show that rice AGO1d, which is specifically expressed in anther wall cells before and during meiosis, associates with both miR2118 and miR2275 to mediate phasiRNA biogenesis. AGO1d preferentially binds to miR2118-triggered 21-nucleotide (nt) phasiRNAs with a 5'-terminal uridine, suggesting a dual role in phasiRNA biogenesis and function. Depletion of AGO1d causes a reduction of 21- and 24-nt phasiRNAs and temperature-sensitive male sterility. At lower temperatures, anthers of the ago1d mutant predominantly show excessive tapetal cells with little starch accumulation during pollen formation, possibly caused by the dysregulation of cell metabolism. These results uncover an essential role of AGO1d in rice anther development at lower temperatures and demonstrate coordinative roles of AGO proteins during reproductive phasiRNA biogenesis and function.

Combined analysis of transcriptome and metabolome reveals that sugar, lipid, and phenylpropane metabolism are essential for male fertility in temperature-induced male sterile rice.

Photoperiod- and thermosensitive genic male sterility (PTGMS) rice is a vital germplasm resource consisting of two-line hybrid rice in which light and temperature strictly control their fertility changes. Variable environmental conditions present huge risks to the two-lines hybrid seed production. Explaining the regulatory mechanism of male fertility in rice PTGMS lines is an essential prerequisite to ensuring food security production. A group of near-isogenic lines (NILs) of a rice PTGMS line unique to this research group was used for this study. These lines have the same genetic background and regulate male fertility by responding to different temperature changes. Transcriptomic analysis revealed that 315 upregulated genes and 391 regulated genes regulated male fertility in response to temperature changes, and differentially expressed genes (DEGs) were mainly characterized in enrichment analysis as having roles in the metabolic pathways of sugar, lipid and phenylpropanoid. Electron microscopy analysis revealed that a lack of starch accumulation in sterile pollen grains induced by high temperature, with an abnormal exine development and a lack of inner pollen grains. Defective processes for sporopollenin synthesis, sporopollenin transport and pollen wall formation in sterile anthers were verified using qPCR. Targeted metabolomics analysis revealed that most lipids (phospholipids, sphingolipids and fatty acids) and flavonoids (flavones and flavanones) were upregulated in fertile anthers and involved in pollen wall development and male fertility formation, while lignin G units and C-type lignin were the major contributors to pollen wall development. The coding genes for trehalose 6-phosphate phosphatase, beta-1,3-glucanase, phospholipase D and 4-coumarate-CoA ligase are considered essential regulators in the process of male fertility formation. In conclusion, our results indicated that the expression of critical genes and accumulation of metabolites in the metabolism of sugar, lipid, and phenylpropanoid are essential for male fertility formation. The results provide new insights for addressing the negative effects of environmental variation on two-line hybrid rice production.

Cia Zeaxanthin Biosynthesis, OsZEP and OsVDE Regulate Striped Leaves Occurring in Response to Deep Transplanting of Rice.

The rice leaf color mutant B03S was previously generated from the photoperiod- and thermo-sensitive genic male sterile (PTGMS) rice line Efeng 1S, of which male sterility manifests by photoperiod and temperature but exhibits mainly temperature-sensitive characteristics. After these plants were deeply transplanted, the new leaves manifested typical zebra stripe patterns. Here, B03S was subjected to deep and shallow transplanting, shading with soil and aluminum foil, and control conditions in situ to determine the cause of the striped-leaf trait. The direct cause of striped leaves is the base of the leaf sheath being under darkness during deep transplanting, of which the critical shading range reached or exceeds 4 cm above the base. Moreover, typical striped leaves were analyzed based on the targeted metabolome method by ultra-performance liquid chromatography/tandem mass spectrometry (UPLC–MS/MS) combined with transcriptome and real-time quantitative PCR (qPCR)-based verification to clarify the metabolic pathways and transcriptional regulation involved. Carotenoids enter the xanthophyll cycle, and the metabolites that differentially accumulate in the striped leaves include zeaxanthin and its derivatives for photooxidative stress protection, driven by the upregulated expression of OsZEP. These findings improve the understanding of the physiological and metabolic mechanisms underlying the leaf color mutation in rice plants, enrich the theoretical foundation of the nonuniform leaf color phenomenon widely found in nature and highlight key advancements concerning rice production involving the transplanting of seedlings or direct broadcasting of seeds.

Mutation of glucose-methanol-choline oxidoreductase leads to thermosensitive genic male sterility in rice and Arabidopsis.

SummaryThermosensitive genic male sterility (TGMS) lines serve as the major genetic resource for two‐line hybrid breeding in rice. However, their unstable sterility under occasional low temperatures in summer highly limits their application. In this study, we identified a novel rice TGMS line, ostms18, of cultivar ZH11 (Oryza sativa ssp. japonica). ostms18 sterility is more stable in summer than the TGMS line carrying the widely used locus tms5 in the ZH11 genetic background, suggesting its potential application for rice breeding. The ostms18 TGMS trait is caused by the point mutation from Gly to Ser in a glucose‐methanol‐choline (GMC) oxidoreductase; knockout of the oxidoreductase was previously reported to cause complete male sterility. Cellular analysis revealed the pollen wall of ostms18 to be defective, leading to aborted pollen under high temperature. Further analysis showed that the tapetal transcription factor OsMS188 directly regulates OsTMS18 for pollen wall formation. Under low temperature, the flawed pollen wall in ostms18 is sufficient to protect its microspore, allowing for development of functional pollen and restoring fertility. We identified the orthologous gene in Arabidopsis. Although mutants for the gene were fertile under normal conditions (24°C), fertility was significantly reduced under high temperature (28°C), exhibiting a TGMS trait. A cellular mechanism integrated with genetic mutations and different plant species for fertility restoration of TGMS lines is proposed.

Influence of Gender Bias on Distribution of Hybrid Sterility in Rice.

Hybrid sterility genes define species identities, setting reproductive barriers between distantly related Oryza relatives. They induce allelic-specific selective gametic abnormalities by killing pollens, embryo sacs, or both, and thus resulting in the male specific transmission ratio distortion (mTRD), female specific transmission ratio distortion (f TRD), and/or sex-independent transmission ratio distortion (siTRD) in hybrids. Although more than 50 hybrid sterility genes have been reported, comprehensive analysis on the distributional pattern of TRD systems in Oryza species is limited. In this review, we surveyed the TRD systems and the underlying possible mechanisms in these species. In rice, pollen killers which cause mTRD are often observed in higher frequency than egg killers and gamete eliminators, which are factors affecting f TRD and siTRD, respectively. Due to the rather massive population of pollen grains, their reduction in the number caused by hybrid sterility possesses a smaller selective disadvantage to the hybrid individuals, in contrast to female gamete abortion. The pattern of TRD distribution displays less abundancy in siTRD. It suggests that fixation of siTRD might require a certain time rather than single sex-specific factors. The presence of linked sterility factors worked for mTRD and f TRD, and strength of their linkage in chromosomal regions might determine the type of sterility and TRD. The study of TRD systems has a potential to reveal the relationships between selfish genes and their functions for reproductive isolation.

Preliminary Research on Intelligent Baking Room Dehydration and Drying Technology for Rice Sterile Seeds

The quality of rice seed is very important for rice production. In the process of rice seed production, seed drying is an important link in the whole process of seed quality control, and the problems in mechanical drying are important factors affecting the quality of rice seed. It is easy to encounter rain weather when the sterile seeds are dried by natural air, which seriously affects the seed vigor. High-quality rice seed is an important guarantee to achieve high yield of rice, and seed germination rate is one of the factors directly affecting rice yield. The drying process has the greatest effect on the quality and germination rate of sterile rice seeds. Selecting reasonable drying technology is an important means to ensure the germination rate of sterile rice seeds. In this paper, the dewatering and drying technology of rice sterile line seeds in intelligent baking room was studied. An adaptive integral sliding mode control algorithm based on Smith prediction was proposed for intelligent baking room temperature. The lag in the system was compensated by Smith prediction; the uncertainty of the system model and the complex interference under variable operating conditions were overcome by the robustness of sliding mode control. And the chattering of the sliding mode was eliminated by the adaptive integral sliding mode surface. The seeds of three rice varieties were dried, the drying dehydration rate, seed germination rate, and seed vigor were measured, and the changes of seed moisture and temperature during drying were observed. The results showed that there was little difference in the seed vigor after drying and air dried, which proved that the dryer could be used to dry different kinds of male sterile seeds. Compared with air dried seeds, the germination rate and germination potential of all mechanically dried seeds have little difference, and the overall difference of germination index and vitality index is also small. This study solved the problem that the seeds of male sterile lines could not be dehydrated and dried in time of rainy weather, which is of great significance to improve seed quality and economic benefits.

Heat Stress During Gametogenesis Irreversibly Damages Female Reproductive Organ in Rice

Heat stress during gametogenesis leads to spikelet sterility. To ascertain the role of female reproductive organ (pistil), two rice genotypes N22 and IR64 with contrasting heat stress responses were exposed to control (30 °C) and heat stress (38 °C and 40 °C) during megasporogenesis. Anatomical observations of ovule revealed greater disappearance of megaspore mother cell and nuclei at early stages, and during later stages mature embryo sac without female germ unit, improper positioning of nuclei, and shrunken embryo sac was observed in the sensitive IR64. Under heat stress, a decrease in sugar and starch, increase in H2O2 and malondialdehyde with lower antioxidant enzyme activities were recorded in pistils of both N22 and IR64. Lower accumulation of TCA cycle metabolites and amino acids were noticed in IR64 pistils under heat stress at gametogenesis, whereas N22 exhibited favorable metabolite profiles. At heading, however, N22 pistils had higher carbohydrate accumulation and better ROS homeostasis, suggesting higher recovery after heat stress exposure. In summary, the results indicate that heat stress during megasporogenesis leads to irreversible anatomical and physiological changes in pistil and alters metabolic signatures leading to increased spikelet sterility in rice. Mechanisms identified for enhanced heat tolerance in pistil can help in developing rice varieties that are better adapted to future hotter climate.

Precise genetic mapping of Rf18(t), a new fertility restorer gene from 'Nipponbare' for wild abortive cytoplasmic male sterility in rice (Oryza sativa L.).

We mapped Rf18(t), a Restorer-of-fertility gene for wild abortive cytoplasmic male sterility from the japonica maintainer 'Nipponbare', to chromosome 1. The best candidate gene, LOC_Os01g71320, is predicted to encode hexokinase. Three-line hybrid rice obtained through cytoplasmic male sterility (CMS) has helped increase the yield of rice globally, and the wild abortive (WA)-type cytoplasm from wild rice (Oryza rufipogon Griff.) is used widely in three-line indica hybrids. The identification and mapping of the Restorer-of-fertility (Rf) genes in maintainer lines aided in uncovering the genetic basis of fertility restoration of WA-type CMS and the development of WA-type hybrids. In this study, we identified a new Rf gene, Rf18(t), for WA-type CMS from the japonica maintainer line 'Nipponbare' using a chromosome segment substitution line population derived from a cross between the indica line 9311 and 'Nipponbare.' Using a substitution mapping strategy, Rf18(t) was delimited to a 48-kb chromosomal region flanked by molecular marker loci ID01M28791 and ID01M28845 on chromosome 1. By comparative sequence analyses, we propose that LOC_Os01g71320 is the most likely candidate gene for Rf18(t), and it is predicted to encode hexokinase. Furthermore, Rf18(t) was found to function in fertility restoration probably by a posttranscriptional mechanism and its function is dependent on the genetic background of 9311. These results broaden our knowledge on the mechanism of fertility restoration of WA-type CMS lines and will facilitate the development of WA-type rice hybrids.

WD40-domain protein GORI is an integrative scaffold that is required for pollen tube growth in rice

ABSTRACT The pollen tube plays a critical role in angiosperm plants by delivering sperm gametes for double fertilization. Although the molecular mechanisms underlying pollen tube germination and growth are crucial to crop plants, they are poorly understood. Here, we describe recent advancements in the understanding of the role of the WD40-domain protein in regulating pollen germination and discuss future directions to investigate its role in rice. GORI encodes a seven-WD40-motif protein that interacts with an AP180 N-terminal homology (ANTH)-domain protein, which modulates clathrin-mediated endocytosis (CME), and regulates Rac6 activity in the apical plasma membrane of elongating pollen tubes. Loss of function of GORI or Rac6 reduces pollen germination and tube growth, thereby resulting in male sterility in rice. In contrast, overexpression of Rac6 increases pollen tube elongation, with this effect being rescued by GORI overexpression. In the absence of ANTH, pollen germination was reduced, similar to the results observed after inhibitor treatment, indicating that pollen germination partially requires CME. Our findings demonstrated that the GORI protein is a positive regulator of pollen germination and tube growth, serving as a link between Rac6 activity regulation and ANTH-mediated endocytosis.

A natural allele of OsMS1 responds to temperature changes and confers thermosensitive genic male sterility

Changes in ambient temperature influence crop fertility and production. Understanding of how crops sense and respond to temperature is thus crucial for sustainable agriculture. The thermosensitive genic male-sterile (TGMS) lines are widely used for hybrid rice breeding and also provide a good system to investigate the mechanisms underlying temperature sensing and responses in crops. Here, we show that OsMS1 is a histone binding protein, and its natural allele OsMS1wenmin1 confers thermosensitive male sterility in rice. OsMS1 is primarily localized in nuclei, while OsMS1wenmin1 is localized in nuclei and cytoplasm. Temperature regulates the abundances of OsMS1 and OsMS1wenmin1 proteins. The high temperature causes more reduction of OsMS1wenmin1 than OsMS1 in nuclei. OsMS1 associates with the transcription factor TDR to regulate expression of downstream genes in a temperature-dependent manner. Thus, our findings uncover a thermosensitive mechanism that could be useful for hybrid crop breeding.

OsSPLs Regulate Male Fertility in Response to Different Temperatures by Flavonoid Biosynthesis and Tapetum PCD in PTGMS Rice.

Photoperiod and thermo-sensitive genic male sterile (PTGMS) rice is an important resource for two line hybrid rice production. The SQUAMOSA–promoter binding, such as the (SPL) gene family, encode the plant specific transcription factors that regulate development and defense responses in plants. However, the reports about SPLs participating in male fertility regulation are limited. Here, we identified 19 OsSPL family members and investigated their involvement in the fertility regulation of the PTGMS rice lines, PA2364S and PA2864S, with different fertility transition temperatures. The results demonstrated that OsSPL2, OsSPL4, OsSPL16 and OsSPL17 affect male fertility in response to temperature changes through the MiR156-SPL module. WGCNA (weighted gene co-expression network analysis) revealed that CHI and APX1 were co-expressed with OsSPL17. Targeted metabolite and flavonoid biosynthetic gene expression analysis revealed that OsSPL17 regulates the expression of flavonoid biosynthesis genes CHI, and the up regulation of flavanones (eriodictvol and naringenin) and flavones (apigenin and luteolin) content contributed to plant fertility. Meanwhile, OsSPL17 negatively regulates APX1 to affect APX (ascorbate peroxidase) activity, thereby regulating ROS (reactive oxygen species) content in the tapetum, controlling the PCD (programmed cell death) process and regulating male fertility in rice. Overall, this report highlights the potential role of OsSPL for the regulation of male fertility in rice and provides a new insight for the further understanding of fertility molecular mechanisms in PTGMS rice.