Search Results (22)

Thermosensitive genic male sterility (TGMS) lines are vital for two-line hybrid breeding. However, the molecular mechanism in the reverse TGMS line 05ms in eggplant remains unclear. Weighted gene co-expression network analysis (WGCNA) of RNA-seq data revealed modules correlated with fertility conversion enriched in carbohydrate metabolism, lipid metabolism, and mRNA surveillance pathways. Hub genes within these modules were predominantly associated with sugar-related processes, fatty acid metabolism, and nucleotide processing. BSA-seq defined candidate genomic intervals. Integrated analysis of BSA-seq intervals and transcriptomic data identified a candidate gene, SmHTH, with consistently lower expression in 05ms compared to S63. Its homologs exhibited temperature-induced expression, possibly compensating for SmHTH deficiency under high temperatures to restore fertility. The homologs co-expressed with three transcription factors are likely intricately linked to this response. We propose a compensatory model demonstrating that low SmHTH expression at low temperatures disrupts key metabolic pathways, causing male sterility. Conversely, elevated expression of homologous genes and transcription factors (TFs) at higher temperatures compensates for the loss of SmHTH function, thereby restoring fertility. The findings of this research not only deepen the theoretical understanding of plant male sterility mechanisms but also provide valuable resources for developing stress-resilient vegetable varieties through modern breeding techniques. Full article

Abiotic stresses increasingly threaten wheat (Triticum aestivum L.) productivity by impairing pollen development and fertilization, yet the molecular regulators that coordinate reproductive success with environmental resilience remain underexplored. Here, we present a comprehensive genome-wide analysis of the Pollen Olea europaea I (POEI) protein family in common wheat. A total of 104 TaPOEI genes were identified and classified into six phylogenetic clades, each sharing conserved exon–intron structures and key protein motifs. Promoter analysis revealed abundant cis-elements associated with phytohormone signaling and abiotic stress responses. Notably, TaPOEI 16-A was preferentially expressed in anthers, showing high expression during early anther development and responding to both high- and low-temperature stresses. Pairwise comparison between thermosensitive genic male-sterile wheat lines and fertile lines suggests a potential role for TaPOEI 16-A in regulating male fertility in response to temperature fluctuations. Our comprehensive analysis establishes a foundation for future functional studies of the TaPOEI family and provides insights into wheat fertility and stress resilience enhancement. Full article

This study aimed to develop an aromatic thermosensitive genic male sterile (TGMS) line in indica rice using CRISPR/Cas9 technology. The TMS5 and FGR in the high-quality conventional rice variety Huahang 48 were targeted for editing using CRISPR/Cas9 technology. CRISPR/Cas9 vectors designed for TMS5 and FGR were constructed and introduced into rice calli through Agrobacterium-mediated transformation. Transgenic seedlings were subsequently regenerated, and the target sites of the edited plants were analyzed via sequencing. A total of fifteen T₀ double mutants were successfully obtained. Three mutants without T-DNA insertion were screened in the T₁ generation by the PCR detection of hygromycin gene fragments, and homozygous mutants without T-DNA insertion were screened in the T₂ generation by the sequencing analysis of the mutation sites, named Huahang 48s. Huahang 48s exhibited complete sterility at 24 °C and pollen transfer at 23 °C. The 2-acetyl-1-pyrroline (2-AP) content was detected in the young panicles, leaves, and stems of Huahang 48s. The leaves of Huahang 48s had the highest 2-AP content, contrasting with the absence of 2-AP in HuaHang 48. F₁ hybrids that crossed Huahang 48s with two high-quality restorer lines were superior to the two parents in terms of yield per plant and 1000-grain weight. Huahang 48s has a certain combining ability and application potential in two-line cross breeding. The successful application of CRISPR/Cas9 technology in Huahang 48 established a foundation for developing aromatic TGMS lines, providing both theoretical insights and practical materials for breeding efforts. Full article

Glume-opening of thermosensitive genic male sterile (TGMS) rice (Oryza sativa L.) lines after anthesis is a serious problem that significantly reduces the yield and quality of hybrid seeds. However, the molecular mechanisms regulating the opening and closing of rice glumes remain largely unclear. In this study, we report the isolation and functional characterization of a glum-opening mutant after anthesis, named gom1. gom1 exhibits dysfunctional lodicules that lead to open glumes following anthesis. Map-based cloning and subsequent complementation tests confirmed that GOM1 encodes a receptor-like kinase (RLK). GOM1 was expressed in nearly all floral tissues, with the highest expression in the lodicule. Loss-of-function of GOM1 resulted in a decrease in the expression of genes related to JA biosynthesis, JA signaling, and sugar transport. Compared with LK638S, the JA content in the gom1 mutant was significantly reduced, while the soluble sugar, sucrose, glucose, and fructose contents were significantly increased in lodicules after anthesis. Together, we speculated that GOM1 regulates carbohydrate transport in lodicules during anthesis through JA and JA signaling, maintaining a higher osmolality in lodicules after anthesis, which leads to glum-opening. Full article

This study aimed to develop and genetically characterize thermosensitive genic male-sterility (TGMS) lines for use in hybrid rice (Oryza sativa L.) breeding. Male-sterile F₂ to F₄ generation lines were screened during the high-temperature summer season, and ratoon crops of selected male-sterile rows were harvested for pure seed. Sixty-six F₅ TGMS lines were genotyped using DNA markers controlling 16 traits from the LSU80 QA/QC Rice PlexSeq SNP Panel. Ten TGMS lines with desirable traits that included semidwarf, glabrous, non-aromatic, long-grain, narrow brown leaf spot resistance, and blast resistance genes were selected for further genotypic characterization using markers for low chalkiness (chalk5), wide compatibility (S5-n), cold tolerance (qSCT-11 and qCST-12), and anaerobic germination (AG1 and AG2). TGMS lines TIL21051S and TIL21052S possess favorable alleles for each of the genes evaluated in this study and are desirable parents for two-line hybrid breeding in the southeast United States. TIL21044S, TIL21095S, TIL21060S, and TIL21066S each contain three blast resistance genes and have potential as parental lines. TIL21014S-2, TIL21015S, and TIL21016S-1 include the fgr allele for aroma and can also be used as parental lines for aromatic two-line hybrids. Full article

Wheat hybrids have been widely demonstrated to have remarkable heterosis or hybrid vigor in increasing yield potential and stability since the 1960s. Two-line hybrid wheat can achieve higher yields than local varieties, especially in marginal environments. However, the commercial application of hybrid wheat is hindered by higher seed costs, primarily due to lower yields in hybrid seed production. Stigma exsertion has been verified as a decisive factor in increasing rice’s hybrid seed yield, but more investigation is needed in hybrid wheat breeding and production. In this study, four thermo-photo-sensitive genic male sterile lines, including K41S, K64S, K66S, and K68S, with different stigma exsertion rates (SERs) were used to compare the differences in floral architecture relative to stigma exsertion over two growing seasons. The results revealed that the K41S and K64S exhibited a relatively higher SER at 21.87% and 22.81%, respectively. No exserted stigma was observed in K66S, and K68S had an SER of only 0.82%. This study found that the stigma length, glume width and the length–width ratio of the glume were significantly correlated with the SER, with correlation coefficients of 0.46, −0.46 and 0.60, respectively. Other stigma features such as the branch angle, stretch width and hairbrush length, as well as the glume length, also had a weakly positive correlation with SER (r = 0.09–0.27). For K41S and K64S, the SER was significantly affected by the differences in the stigma branch angle and stigma stretch width among florets. A cross-pollination survey showed that the out-crossing ability of florets with an exserted stigma was about three times as high as that of florets with a non-exserted stigma. As a result, the stigma-exserted florets that accounted for 21.87% and 22.81% of the total florets in K41S and K64S produced 46.80% and 48.53% of the total cross-pollinated seeds in both sterile lines. These findings suggest that a longer stigma combined with a slender glume appears to be the essential floral feature of stigma exsertion in sterile wheat lines. It is expected that breeding and utilizing sterile lines with a higher SER would be a promising solution to cost-effective hybrid wheat seed production. Full article

Rice B03S mutants with intermittent leaf discoloration were developed from the photoperiod- and thermosensitive genic male sterile (PTGMS) rice line Efeng 1S. After these plants were deeply transplanted, the new leaves manifested typical stripe patterns. In this study, deep and shallow transplantation of B03S was carried out, and aluminum shading was performed directly on the leaf sheath. It was determined that the reason for the appearance of the striped leaf trait was that the base of leaf sheath lacked light, at which time the sheath transformed from the source organ to the sink organ in rice. To elucidate the related metabolic changes in glycometabolism and abscisic acid (ABA) biosynthesis and transcriptional regulation in the leaf sheath, ultra-performance liquid chromatography/tandem mass spectrometry (UPLC-MS/MS) combined with transcriptome and real-time quantitative PCR (qPCR) validation were used for analysis after deep and shallow transplantation. The result indicates that the leaf sheath may need to compete with the new leaves for sucrose produced by the photosynthesis of old leaves in response to lacking light at the base of sheath. Moreover, the ABA content increases in the leaf sheath when the gene expression of ABA2 and AAO1 is upregulated at the same time, enhancing the plant’s resistance to the adverse condition of shading at the leaf sheath. Furthermore, exogenous spraying of B03S with ABA solution was carried out to help recovery under shading stress. The result indicates that the synthesis of endogenous ABA in the leaf sheath is reduced by spraying ABA. At the same time, ABA regulates sucrose metabolism by inhibiting the expression of the SUS gene. This allows for more sucrose synthesized by the old leaves to be transported to the new leaves, resulting an obvious recovery effect of the strip leaf character due to the re-balance of sugar supply and demand in B03S. These findings improve the understanding of the physiological function and metabolic mechanism of the rice leaf sheath, provide a theoretical basis for uneven leaf coloration in nature, and provide theoretical guidance for rice production via seedling transplantation or direct seeding. Full article

Seed storability has a significant impact on seed vitality and is a crucial genetic factor in maintaining seed value during storage. In this study, RNA sequencing was used to analyze the seed transcriptomes of two rice thermo-sensitive genic male sterile (TGMS) lines, S1146S (storage-tolerant) and SD26S (storage-susceptible), with 0 and 7 days of artificial accelerated aging treatment. In total, 2658 and 1523 differentially expressed genes (DEGs) were identified in S1146S and SD26S, respectively. Among these DEGs, 729 (G1) exhibited similar regulation patterns in both lines, while 1924 DEGs (G2) were specific to S1146S, 789 DEGs (G3) were specific to SD26S, and 5 DEGs (G4) were specific to contrary differential expression levels. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that “translation”, “ribosome”, “oxidative phosphorylation”, “ATP-dependent activity”, “intracellular protein transport”, and “regulation of DNA-templated transcription” were significantly enriched during seed aging. Several genes, like Os01g0971400, Os01g0937200, Os03g0276500, Os05g0328632, and Os07g0214300, associated with seed storability were identified in G4. Core genes Os03g0100100 (OsPMEI12), Os03g0320900 (V2), Os02g0494000, Os02g0152800, and Os03g0710500 (OsBiP2) were identified in protein–protein interaction (PPI) networks. Seed vitality genes, MKKK62 (Os01g0699600), OsFbx352 (Os10g0127900), FSE6 (Os05g0540000), and RAmy3E (Os08g0473600), related to seed storability were identified. Overall, these results provide novel perspectives for studying the molecular response and related genes of different-storability rice TGMS lines under artificial aging conditions. They also provide new ideas for studying the storability of hybrid rice. Full article

Rice blast caused by Magnaporthe oryzae is one of the most serious rice diseases worldwide. The early indica rice thermosensitive genic male sterile (TGMS) line HD9802S has the characteristics of stable fertility, reproducibility, a high outcrossing rate, excellent rice quality, and strong combining ability. However, this line exhibits poor blast resistance and is highly susceptible to leaf and neck blasts. In this study, backcross introduction, molecular marker-assisted selection, gene chipping, anther culture, and resistance identification in the field were used to introduce the broad-spectrum blast-resistance gene R6 into HD9802S to improve its rice blast resistance. Six induction media were prepared by varying the content of each component in the culture medium. Murashige and Skoog’s medium with 3 mg/L 2,4-dichlorophenoxyacetic acid, 2 mg/L 1-naphthaleneacetic acid, and 1 mg/L kinetin and N6 medium with 800 mg/L casein hydrolysate, 600 mg/L proline, and 500 mg/L glutamine could improve the callus induction rate and have a higher green seedling rate and a lower white seedling rate. Compared to HD9802S, two doubled haploid lines containing R6 with stable fertility showed significantly enhanced resistance to rice blast and no significant difference in spikelet number per panicle, 1000-grain weight, or grain shape. Our findings highlight a rapid and effective method for improving rice blast resistance in TGMS lines. Full article

Rice blast caused by pathogenic fungus Magnaporthe oryzae is one of the most serious diseases in rice. The pyramiding of effective resistance genes into rice varieties is a potential approach to reduce the damage of blast disease. In this study, combinations of three resistance genes, Pigm, Pi48 and Pi49, were introduced into a thermo-sensitive genic male sterile (PTGMS) line Chuang5S through marker-assisted selection. The results showed that the blast resistance of improved lines increased significantly compared with Chuang5S, and the three gene pyramiding lines (Pigm + Pi48 + Pi49) had higher rice blast resistance level than monogenic line and digenic lines (Pigm +Pi48, Pigm + Pi49). The genetic backgrounds of the improved lines were highly similar (>90%) to the recurrent parent Chuang5S by using the RICE10K SNP chip. In addition, agronomic traits evaluation also showed pyramiding lines with two or three genes similar to Chuang5S. The yields of the hybrids developed from improved PTGMS lines and Chuang5S are not significantly different. The newly developed PTGMS lines can be practically used for the breeding of parental lines and hybrid varieties with broad spectrum blast resistance. Full article

Two-line hybrid rice production requires environmental genetic male-sterile (EGMS) lines that become sterile in certain environmental conditions. In this study, we aimed to identify the genetic type(s), sterility thresholds, optimum planting date, and efficient seeding density of eight male-sterile lines, including four lines developed at the University of Arkansas, Rice Research and Extension Center (RREC) and four at the Dale Bumpers National Rice Research Center (DBNRRC) both located in Stuttgart, AR, USA. The lines were screened in growth chambers using different temperatures and day lengths to identify the type of EGMS line and sterility thresholds. A single location was used in testing the first year of the study and three locations used in the second year of the planting date study. All experiments were arranged in randomized blocks with three replications to identify the optimum planting date and seeding rate. Three planting dates were tested each year for the planting date experiment, and three seeding densities were used in the seed density study. The growth chamber study showed that all RREC lines were thermosensitive genic male-sterile lines, and the sterility threshold temperature for one of these lines was >32.2 °C while that of others was >29.4 °C. The type of sterility and threshold of DBNRRC lines were inconclusive due to their genetic heterogenicity and environmental response inconsistency. In Arkansas, April 25 was the optimum planting date for sterile conditions, and a 431 seed/m² seeding rate was required to produce an adequate number of panicles for hybrid seed production while avoiding wasting expensive male-sterile seeds. Full article

Anther dehiscence is an important process to release pollen and then is a critical event in pollination. In the wheat photo-thermo-sensitive genic male sterility (PTGMS) line, pollen cannot release from anther since the anther cannot dehisce during anther dehiscence stage in a sterile condition. In this study, we carried out RNA-sequencing to analyze the transcriptome of one wheat PTGMS line BS366 during anther dehiscence under fertile and sterile conditions to explore the mechanism. We identified 6306 differentially expressed genes (DEGs). Weighted gene co-expression network analysis (WGCNA) and KEGG analysis showed that DEGs were mainly related to “hormone signal transduction pathway” and “starch and sucrose metabolism”. We identified 35 and 23 DEGs related hormone signal transduction and sucrose metabolism, respectively. Compared with conventional wheat Jing411, there were some changes in the contents of hormones, including JA, IAA, BR, ABA and GA3, and sucrose, during three anther dehiscence stages in the sterile condition in BS366. We performed qRT-PCR to verify the expression levels of some critical DEGs of the hormone signaling pathway and the starch and sucrose metabolism pathway. The results showed disparate expression patterns of the critical DEGs of the hormone signaling pathway and the starch and sucrose metabolism pathway in different conditions, suggesting these genes may be involved in the regulation of the anther dehiscence in BS366. Finally, we conducted a hypothesis model to reveal the regulation pathway of hormones and sucrose on anther dehiscence. The information provided new clues to the molecular mechanisms of anther dehiscence in wheat and improved wheat hybrid breeding. Full article

The development of thermosensitive genic male sterile (TGMS) lines is the key to breeding two-line hybrid rice, which has been widely applied in China to increase grain yield. CRISPR/Cas9 has been widely used in genome editing to create novel mutants in rice. In the present study, a super grain quality line, GXU 47, was used to generate a new TGMS line with specific mutations in a major TGMS gene tms5 generated with CRISPR/Cas9-mediated genome editing in order to improve the rice quality of two-line hybrids. A mutagenesis efficiency level of 75% was achieved, and three homozygous T-DNA-free mutant lines were screened out. The mutants exhibited excellent thermosensitive male fertility transformation characteristics with complete male sterility at ≥24 °C and desirable male fertility at around 21 °C. Proteomic analysis based on isobaric tags for relative and absolute quantification (iTRAQ) was performed to unveil the subsequent proteomic changes. A total of 192 differentially expressed proteins (DEPs), including 35 upregulated and 157 downregulated, were found. Gene ontology (GO) analysis revealed that the DEPs were involved in a single-organism biosynthetic process, a single-organism metabolic process, oxidoreductase activity, and catalytic activity. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that the DEPs were involved in ubiquinone and other terpenoid quinone biosynthesis, the biosynthesis of secondary metabolites, metabolic pathways, and phenylpropanoid biosynthesis. Our study shows that high mutation efficiency was achieved in both target sites, and T-DNA-free mutant lines were obtained in the T₁ generation. The present study results prove that it is feasible and efficient to generate an excellent mutant line with CRISPR/Cas9, which provides a novel molecular mechanism of male sterility caused by the mutation of tms5. Full article

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. Full article

Zhu1S is a thermosensitive genic male-sterile (TGMS) line of rice possessing outstanding combining ability and low critical temperature, which has been extensively utilized as a female parent in two-line hybrid ricebreeding. However, the fertility of Zhu 1S during hybrid seed production is frequently affected by high temperature, thus leading to its fertility alteration and aborted hybrid seed production. To understand its fertility conversion mechanism under high temperature, we employed transcriptomics analyses on the anthers of young panicles of Zhu 1S during the fertility alternation sensitivity stage under high (Zhu 1S-H) and low (Zhu 1S-L) temperatures. The results showed that a total of 1119 differentially expressed genes (DEGs) were identified between Zhu 1S-H and Zhu1S-L anthers, including 680 up-regulated and 439 down-regulated genes. Bioinformatics analysis of these DEGs revealed that the high temperature induction caused fertility-sterility conversion in Zhu1S, mainly by decreasing the mRNA abundances of important genes closely related to plant hormone and MAPK signal pathway and transcriptional regulation factors, thereby impeding the growth and development of the anther of Zhu 1S, which ultimately affected the fertility transition of Zhu 1S under high temperature. The protein–protein interaction network analysis indicates that transcription factor OsTIFY11C possibility plays a central role in the fertility transition of Zhu 1S under high temperature.The present studies offer a theoretical foundation for further research into the molecular mechanism underlying fertility conversion in TGMS line Zhu 1S. Full article