Male Sterility Research Articles

Progress and prospects in the molecular basic research on important traits in sorghum

Sorghum (Sorghum bicolor) is a significant crop serving food, energy, feed, and industrial raw materials, featuring extensive growth adaptability and diverse utility values. Despite the achievements in sorghum breeding in the last decades, conventional breeding methods still confront challenges such as lengthy breeding cycles, low efficiency, and complex genetic backgrounds. With the rapid advancement of molecular biology, genetics, and bioinformatics, molecular breeding has carved new pathways for enhancing sorghum yield and quality. This article reviews the molecular basic research progress in the key agronomic and adaptive traits of sorghum, including grain yield, grain quality, flowering time, plant height, tillering, stress resistance, and male sterility, and discusses future research priorities, offering novel insights and approaches for sorghum breeding.

The Residual Activity of Fatty Acyl-CoA Reductase Underlies Thermo-Sensitive Genic Male Sterility in Rice.

Photoperiod/thermo-sensitive genic male sterility (P/TGMS) is critical for rice two-line hybrid system. Previous studies showed that slow development of pollen is a general mechanism for sterility-to-fertility conversion of TGMS in Arabidopsis. However, whether this mechanism still exists in rice is unknown. Here, we identified a novel rice TGMS line, ostms16, which exhibits abnormal pollen exine under high temperature and fertility restoration under low temperature. In mutant, a single base mutation of OsTMS16, a fatty acyl-CoA reductase (FAR), reduced its enzyme activity, leading to defective pollen wall. Under high temperature, the mOsTMS16M549I couldn't provide sufficient protection for the microspores. Under low temperature, the enzyme activity of mOsTMS16M549I is closer to that of OsTMS16, so that the imperfect exine could still protect microspore development. These results indicated whether the residual enzyme activity in mutant could meet the requirement in different temperature is a determinant factor for fertility conversion of P/TGMS lines. Additionally, we previously found that res2, the mutant of a polygalacturonase for tetrad pectin wall degradation, restored multiple TGMS lines in Arabidopsis. In this study, we proved that the osres2 in rice restored the fertility of ostms16, indicating the slow development is also suitable for the fertility restoration in rice.

Residual Longevity of Recaptured Sterile Mosquitoes as a Tool to Understand Field Performance and Reveal Quality

Invasive mosquito species, such as Aedes albopictus, pose significant threats to both ecosystems and public health due to their role in transmitting diseases, such as dengue, Zika, and chikungunya. The Sterile Insect Technique (SIT) is a promising vector control strategy aimed at reducing mosquito populations by releasing sterile males to mate with wild females and reduce their reproduction rates. In this study, we employed the captive cohort method, which assesses the remaining longevity of randomly caught released individuals, to assess the longevity and frailty dynamics of sterile and non-sterile Ae. albopictus males. Using a mark–release–recapture approach (MRR), we compared the residual lifespan of sterile and non-sterile released males with that of wild, non-sterile males, aiming to understand the frailty dynamics of released males and, therefore, their quality and field performance. Contrary to expectations, our results revealed that released sterile males showed increased longevity compared to non-sterile males. Further, the marking process did not impact the longevity between lab-kept and marked males, suggesting that the marking process does not adversely affect survival under controlled conditions. These findings underscore the importance of optimizing pre-release and mass-rearing practices to enhance the effectiveness of SIT programs. Our study also demonstrates for the first time the use of the captive cohort method for understanding the biological dynamics of sterile mosquito populations in SIT programs, providing valuable insights for improving vector control strategies.

EMBRYONIC FLOWER 1 regulates male reproduction by repressing the jasmonate pathway downstream transcription factor MYB26.

The evolutionarily conserved Polycomb repressive complexes (PRC) mediate genome-wide transcriptional silencing and regulate a plethora of development, as well as environmental responses in multicellular organisms. The PRC2-catalyzed trimethylation of lysine 27 on histone H3 (H3K27me3) is recognized by reader-effector modules of PRC1 to implement gene repression. Here, we report that the Arabidopsis (Arabidopsis thaliana) H3K27me3 effector EMBRYONIC FLOWER 1 (EMF1) interacts with and constrains the R2R3 DNA binding transcription factor MYB26 by a eudicot-conserved motif in the stamen. MYB26 activates the transcription of two NAC domain genes, NAC SECONDARY WALL THICKENING PROMOTING FACTOR1 (NST1) and NST2, whose encoded proteins mediate anther secondary cell thickening in jasmonate (JA)-regulated stamen maturation. In this process, the transcriptional activity of MYB26 is negatively modulated by the JAZ-PRC repressive complex to precisely regulate the expression of NST1 and NST2. Disruption of EMF1 repression stimulates MYB26, leading to the excessive transcription of the two NAC genes and male sterility. Our results reveal a novel mechanism in polycomb-mediated gene silencing and illustrate that the plant Polycomb complex regulates stamen development by preventing the hypersensitivity of JA responses in male reproduction.

Genetic Analysis of Combining Ability and Gene Action in Sunflower (Helianthus annuus L.) Hybrids through Line × Tester Design

This study was conducted to evaluate the combining ability of six cytoplasmic male sterile (CMS) lines and nine testers using the line × tester method. Biometrical data were recorded for ten agronomic traits: days to 50% flowering, days to maturity, plant height, head diameter, seed filling percentage, 100-seed weight, volume weight, hull content, oil content, and seed yield per plant. The results indicated that specific combining ability (SCA) variance was higher than general combining ability (GCA) variance for most traits, except for days to 50 percent flowering and seed filling percentage, suggesting that non-additive gene action predominantly governed the inheritance of these traits. The GCA of parents did not always align with their per se performance, indicating that GCA should be the primary criterion for selecting parents in breeding programs. Three CMS lines, CMS-103A, CMS-10A and CMS-112A, along with testers EC-75717, EC-502036, EC-279309-1 and R-271-1, were identified as good general combiners for seed yield and yield-contributing traits. Among the hybrids, CMS-243A × EC-279309-1, CMS-243A × EC-75717 and CMS-103A × EC-75717 exhibited the highest SCA effects for seed yield per plant. These findings underscore the importance of selecting superior general combiners for traits like oil content and seed yield to develop high-yielding sunflower hybrids.

Gene expression profiles and metabolic pathways responsible for male sterility in cybrid pummelo.

Abnormal expression of genes regulating anther and pollen development and insufficient accumulation of male sterility (MS)- related metabolites lead to MS in cybrid pummelo Male sterility (MS) is a major cause of seedlessness in citrus, which is an important trait for fresh fruit. Understanding the mechanism of MS is important for breeding seedless citrus cultivars. In this study, we dissected the transcriptional, metabolic and physiological mechanisms of MS in somatic cybrid of pummelo (G1 + HBP). G1 + HBP exhibited severe male sterility, manifesting as retarded anther differentiation, abnormal anther wall development (especially tapetum and endothecium), and deficient pollen wall formation. In the anthers of G1 + HBP, the expression of genes regulating anther differentiation and tapetum development was abnormal, and the expression of genes regulating endothecium secondary lignification thickening and pollen wall formation was down-regulated. The transcription of genes involved in MS-related biological processes, such as jasmonic acid (JA) signaling pathway, primary metabolism, flavonoid metabolism, and programmed cell death, was altered in G1 + HBP anthers, and the accumulation of MS-associated metabolites, including fatty acids, amino acids, sugars, ATP, flavonols and reactive oxygen species (ROS), was down-regulated in G1 + HBP anthers. In summary, abnormal expression of key genes regulating anther and pollen development, altered transcription of key genes involved in MS-related metabolic pathways, and insufficient accumulation of MS-related metabolites together lead to MS in G1 + HBP. The critical genes and the metabolism pathways identified herein provide new insights into the formation mechanism of MS in citrus and candidate genes for breeding seedless citrus.

The evolution and maintenance of trioecy with cytoplasmic male sterility.

Trioecy, the co-existence of females, males and hermaphrodites, is a rare sexual system in plants that may be an intermediate state in transitions between hermaphroditism and dioecy. Previous models have identified pollen limitation as a necessary condition for the evolution of trioecy from hermaphroditism. In these models, the seed-production and pollen production of females and males relative to those of hermaphrodites, respectively, are compromised by self-fertilization by hermaphrodites under pollen- limitation. Here, we investigate the evolution of trioecy via the invasion of cytoplasmic male sterility (CMS) into androdioecious populations in which hermaphrodites co-occur with males and where the male determiner is linked to a (partial) fertility restorer. We show that the presence of males in a population renders invasion by CMS more difficult. However, the presence of males also facilitates the maintenance of trioecy even in the absence of pollen limitation by negative frequency-dependent selection, because males reduce the transmission of CMS by females by siring sons (which cannot transmit CMS). We discuss our results in light of empirical observations of trioecy in plants and its potential role in the evolution of dioecy.

Improving the green mass quality with the help of heterosis breeding

The most reliable and effective way to improve both productivity and quality of green mass is the use of heterosis breeding. The purpose of the current work was to study the heterosis effect of green mass quality indicators in sorghum-Sudan hybrids developed based on cytoplasmic male sterility. The object of the study was the sorghum-Sudan hybrids developed by the Agricultural Research Center “Donskoy”. Dry matter productivity varied among the hybrids within 746-1202 g/m2 . The highest values have been observed in the combinations with the CMS-line ‘APV-1115’ (1045 g/m2 ). The heterosis effect on the dry matter productivity was found in all hybrids. Four hybrids ‘APV-1115 x Kudesnitsa’, ‘APV-1115 x ChT-22’, ‘Dzhetta x ChT-2’, ‘APV-1115 x Svetloplenchataya 4’ have significantly exceeded the standard in terms of dry matter productivity by 184-270 g/m2 with high rates of hypothetical (38.1-64.1%), true (93.2-142.0%) and competitive (19.8-28.9%) heterosis. Digestible protein yield has ranged from 49 to 84 g/m2 . Among the hybrids, the greatest heterosis effect was identified in the hybrids ‘Demetra x Svetloplenchataya 4’ (Hhyp.=43.4%, Htrue.=125.4%), ‘Dzhetta x Yaktik’ (Hhyp.=70.5%, Htrue.=142.3%), ‘APV-1115 x Kudestitsa’ (Hhyp.=72.8%, Htrue.=108.9%), ‘APV-1115 x Svetloplenchataya 2’ (Hhyp.=81.0%, Htrue.=118.4%), ‘APV1115 x ChT-22’ (Hhyp.=103.3%, Htrue.=118.4%). In the course of the study, there has been found that according to the content of crude protein, ash, oil, fiber and nitrogen-free extractive substances (NES), the most hybrids possessed partial or incomplete dominance of large values. However, to obtain even a slight heterosis according to the content of protein, fiber, and NES content, at least one parent had to have high trait values. Based on the complex of the heterotic effect according to the green mass quality indicators, there have been identified such hybrids as ‘Dzhetta x Yaktik’, which exceeded the large parental form in terms of dry matter (Htrue.=10.5%) and oil (Htrue.=14.4%); ‘Dzhetta x ChSS’ which exceeded the large parental form in terms of dry matter and crude protein (Htrue.=14.3 and 1.3%); ‘Demetra x Svetloplenchataya 4’ which exceeded the large parental form in terms of crude protein, dry matter, and ash (Htrue.=3.8; 5.9 and 6.4%, respectively). In general, the data obtained have indicated the prospects of using cytoplasmic male sterility in breeding not only for productivity, but also for quality.

A history of susceptibility to parasites and divergence in solitary, gregarious and agonistic behaviors of embiopterans.

Two species of Haploembia Ramburi (Oligotomidae: Embioptera), nonnative detritivores found in the western USA, display solitary tendencies, not typical for webspinners that usually share silk galleries. Reports from the 1960s based on native populations in Italy highlighted the impact of a gregarine that depressed male sterility and female survivorship in Haploembia solieri (Rambur). Sympatric asexual Haploembia tarsalis (Ross) lives a normal lifespan when parasitized, albeit suffering from reduced fecundity. Our goal is to characterize behavioral repertoires as individuals interact and to develop methods for future investigations focused on the impact of the little-known parasite. We quantified individual responses to conspecifics or other species in 10-min dyadic interactions and, in a 3-day trial, determined whether they aggregated when given dispersed resources. Replicated groups of four adult female H. solieri or H. tarsalis settled away from each other over the 3-day trials. In 10-min bouts of same or different species pairs, focal insects bolted back, retreated and attempted to escape when they encountered one another, especially when the opponent was H. tarsalis. Males courted conspecific females, but were dramatically repelled by H. tarsalis. Serving as a positive control, Oligotoma nigra (Hagen) (Oligotomidae) adult females paired with conspecifics displayed typical webspinner behaviors by sitting together, sharing silk. Haploembia solieri males did not respond negatively to O. nigra, not known to be parasitized by the gregarine, but did so when paired with H. tarsalis. Results align with the prediction that susceptibility to parasitism may have led to antisocial behaviors observed in two Haploembia species.

CRISPR/Cas9 and Anther Culture for Precision Double Haploid Line Production in Controlled Environments

ABSTRACTThe development of mapping populations and quantitative trait loci (QTL) analysis face constraints, in crops exhibiting male sterility and self‐incompatibility under field conditions. Addressing these challenges requires the integration of advanced techniques, including the temporal alteration or excision of centromere histone H3 (CENH3) protein and the use of gene editing tools such as MATRILINEAL (MTL) knockout. Specifically, this can be achieved through Cas9/gRNA‐mediated mutagenesis or Cas9/gRNA‐driven promoter expression systems. These technologies offer efficient means to advance mapping populations and QTL analysis in male sterile and self‐incompatible crops within controlled ecosystems. The doubled haploid (DH) mapping population, traditionally requiring 3 years of generation time via anther culture method, can now be expedited to 2–3 years of generation time using gene editing techniques within controlled environmental systems. Notably, DH mapping populations can be efficiently generated in various crops, including rice, wheat, maize, barley and oats by leveraging gene editing tools. Among these tools, the novel approach of CENH3 protein temporal alteration/excision emerges as highly efficient compared to MTL knockout using Cas9/gRNA‐mediated mutation or Cas9/gRNA promoter expression. However, further investigation is warranted to optimise the regeneration of double haploid populations and enhance QTL analysis in male sterile and self‐incompatible crops under controlled systems.

The effect of T. aestivum chromosomes 1A and 1D on fertility of alloplasmic recombinant (H. vulgare)-T. aestivum lines depending on cytonuclear compatibility.

The effect of T. aestivum L. chromosomes 1A and 1D on fertility of recombinant bread wheat allolines of the same origin carrying the cytoplasm of barley H. vulgare L. and different levels of cytonuclear compatibility was studied. Alloline L-56 included mainly fully sterile (FS) and partially sterile (PS) plants, alloline L-57 included partially fertile (PF) plants and line L-58 included fertile (F) ones. Analysis of morphobiological traits and pollen painting indicated complete or partial male sterility in plants of allolines L-56 and L-57. To differentiate genotypes with cytonuclear coadaptation and genotypes with cytonuclear incompatibility, PCR analysis of the 18S/5S mitochondrial (mt) repeat was performed. Heteroplasmy (simultaneous presence of barley and wheat mtDNA copies) was found in FS, PS, PF and some F plants, which was associated with a violation of cytonuclear compatibility. Wheat-type homoplasmy (hm) was detected in the majority of the fertile plants, which was associated with cytonuclear coadaptation. The allolines used as maternal genotypes were crossed with wheat-rye substitution lines 1R(1A) and 1R(1D). In F1, all plants of PF×1R(1A) and PF×1R(1D) combinations were fertile, and in F2, a segregation close to 3 (fertile) : 1 (sterile) was observed. These results showed for the first time that chromosomes 1A and 1D carry one dominant Rf gene, which controls the restoration of male fertility of bread wheat carrying the cytoplasm of H. vulgare. All plants of F1 combinations FS×1R(1A), FS×1R(1D), PS×1R(1A), PS×1R(1D) were sterile, which indicates that a single dose of genes localized on wheat chromosomes 1A or 1D is not enough to restore male fertility in FS and PS plants. All plants of hybrid combinations F(hm)×1R(1A) and F(hm)×1R(1D) in both F1 and F2 were fertile, that is, fertility of allolines with cytonuclear coadaptation does not depend on wheat chromosomes 1A and 1D.

Sharp decline in male fertility in F2 hybrids of the female-heterogametic silk moth Bombyx.

Sexual selection drives rapid evolution of morphological, physiological, and behavioral traits, especially in males, and it may also drive the rapid evolution of hybrid male sterility. Indeed, the faster male theory of speciation was once viewed as a major cause of Haldane's rule in male-heterogametic XY taxa, but is increasingly being replaced by the genetic conflict hypothesis partly because it cannot explain the faster evolution of hybrid female sterility in female-heterogametic ZW taxa. The theory nonetheless predicts that there should be more genes for hybrid male sterility than for hybrid female sterility even in such taxa, but this remains untested. Thus, finding evidence for the faster male theory of reproductive isolation beyond the F1 generation in ZW systems still represents a challenge to studying the impact of sexual selection. In this study, we examined F2 hybrids between the domesticated silkworm Bombyx mori and the wild silk moth Bombyx mandarina, which have ZW sex determination. We found that although only females showed reduced fertility in the F1 generation, the F2 hybrid males had a significant reduction in fertility compared with the parental and F1 males. Importantly, 27% of the F2 males and 15% of the F2 females were completely sterile, suggesting the presence of recessive incompatibilities causing male sterility in female-heterogametic taxa.

Vasectomy provider decision-making balancing autonomy and non-maleficence: qualitative interviews with providers

Background Male sterilization, or vasectomy, is 99.9% effective at preventing pregnancy with less than a 2% risk of complications. Despite the high efficacy, low risk, low cost, and gender equity benefits of vasectomy, just 2% of women reported that they and their partners relied on vasectomy as their contraceptive method globally in 2019. Health care providers can be both a facilitator and a barrier in men’s health generally, and may be in vasectomy provision as well. This study sought to describe the decision-making rationales of experienced vasectomy providers when evaluating patient candidacy in complex cases. Methods Fifteen vasectomy providers belonging to the global Vasectomy Network Google Group from seven countries participated in online interviews using a semi-structured in-depth interview guide. Providers were asked about their vasectomy training, their reasons for vasectomy provision, challenging cases they have faced, and approaches used to manage challenging cases. Vignettes were used to further elicit decision-making rationale. Thematic analysis was conducted using MAXQDA20. Results Provider decision-making was predicated on ensuring patients were well-informed, able to consent, and certain about their choice to have a vasectomy. Once those foundational conditions were met, providers filtered patient characteristics through their training, laws and policies, sociocultural norms, experience, and peer influence to produce a cost-benefit breakdown. Based on the cost-benefit analysis, providers determined whether to weigh autonomy or non-maleficence more heavily when determining vasectomy patient candidacy. Conclusions Despite clinical best practices that promote prioritizing patient autonomy over non-maleficence, some providers continued to weigh non-maleficence over autonomy in vasectomy patient candidacy evaluations. Non-maleficence was particularly prioritized in cases providers deemed to be at higher risk of regret. The findings of this study suggest vasectomy provider training should emphasize evidence-based best practices in shared decision-making and patient-centered care to facilitate vasectomy provision that honors patient autonomy and rights.

Heat Stress Inhibits Pollen Development by Degrading mRNA Capping Enzyme ARCP1 and ARCP2.

Pollen development and germination are critical for successful generation of offspring in plants, yet they are highly susceptible to heat stress (HS). However, the molecular mechanism underlying this process has not been fully elucidated. In this study, we highlight the essential roles of two mRNA capping enzymes, named Arabidopsis mRNA capping phosphatase (ARCP) 1 and 2, in regulating male and female gamete development. The transmission efficiencies of gametes carrying arcp1 arcp2 from arcp1+/- arcp2-/- and arcp1-/- arcp2+/- mutants are 30% and zero, respectively. These mutants exhibited a significant increase in misshaped pollen, with germination rates approximately half of those in wild type. ARCP1/2 exhibit RNA triphosphatase and RNA guanylyltransferase activities, which are required for proper pollen development. Through RNA-seq analysis, genes involved in pollen development/germination and HS response were identified as downregulated genes in pollen from arcp1+/- arcp2-/- mutant. Furthermore, ARCP2 protein is degraded under HS condition, and inducing the expression of ARCP2 can increase the pollen germination rate under elevated temperature. We propose that HS triggers the degradation of mRNA capping enzymes, which in turn disrupts the transcriptome that required for pollen development and pollen germination and ultimately leads to male sterility.

Fertility restorer gene CaRf and PepperSNP50K provide a promising breeding system for hybrid pepper.

Cytoplasmic male sterility (CMS) is pivotal in plant breeding and widely employed in various crop hybrids, including pepper. However, the functional validation of the restorer of fertility (Rf) gene in pepper has been lacking until now. This study identifies and characterizes CaRf, a single dominant locus crucial for restoring CMS in the pepper strong recovery inbred line Zhangshugang. The CaRf gene encodes a mitochondria-targeted pentatricopeptide repeat protein, validated through the induction of male sterility upon its silencing in hybrid F1 plants. To enhance pepper breeding efficiency, 176 important pepper breeding parent materials were resequenced, and a PepperSNP50K liquid-phase breeding chip was developed, comprising 51 172 markers. Integration of CaRf functional characterization and PepperSNP50K facilitated the development of a high-quality red pepper hybrid. These findings provide significant insights and practical strategies for advancing molecular-designed breeding in peppers.

Disruption of Energy Metabolism and Reactive Oxygen Species Homeostasis in Honglian Type-Cytoplasmic Male Sterility (HL-CMS) Rice Pollen

Honglian type-cytoplasmic male sterility (HL-CMS) is caused by the inter-communication between the nucleus and mitochondria. However, the mechanisms by which sterility genes regulate metabolic alterations and changes in mitochondrial morphology in the pollen of HL-CMS remains unclear. In this study, we compared the morphological differences between the pollen of the male sterile line YA and the near-isogenic line NIL-Rf6 using hematoxylin and eosin staining as well as 4ʹ,6-diamidino- 2-phenylindole (DAPI) staining. HL-CMS is characterized by gametophytic sterility, where the aborted pollen grains are empty and the tapetal layer remains intact. Transmission electron microscopy was employed to observe mitochondrial morphological changes at the microspore stage, revealing that significant mitochondrial alternations, characterized by the formation of 'large spherical mitochondria', occurred at the binucleate stage in the YA line. Additionally, metabolomics analysis revealed decreased levels of metabolites associated with the carbohydrate and flavonoid pathways. Notably, the decrease in flavonoids was found to contribute to an elevation in reactive oxygen species (ROS) levels. Therefore, we propose a model in which rice fertility is modulated by the levels of pollen carbohydrates and flavonoid metabolites, with impaired mitochondrial energy production and reduced flavonoid biosynthesis being the main causes of ROS accumulation and pollen abortion in rice.

Identification of stable restorers and high-yielding hybrids using diverse sorghum male sterile cytoplasmic sources and established pollen parents under different water regimes

Sorghum hybrids demonstrated increased productivity and helped offset the decreasing cultivated areas, particularly in Asia. The diversity in cytoplasmic male sterility systems, stability of restorers and high yield of sorghum is an important factor for achieving food security and sustainability. In sorghum, hybrid production has been limited to A1 cytoplasmic source to date, primarily due to limited number of restorers on other cytoplasmic sources. This work aimed at filling this gap through diversifying and assessing the producibility and stability of cytoplasmic backgrounds across environments, evaluating the stability of restorers across cytoplasms and environments, and evaluating the performance of the hybrid F1 products under different water regimes across cropping seasons. The effects of genotypes and interactions between the genotypes and the tested environments were shown visually using GGE biplot, which also categorized the genotypes based on performance per se and stability. Analysis of variance revealed significant (P< 0.001) differences between genotypes, environments and their interaction components for all the plant characteristics studied. The various GGE biplot patterns revealed that the restorers G4 (IS 33844-5), G5 (M 35-1-19), G8 (ICSV 15014), and G10 (ICSR 13009) were stable across environments and cytoplasms. We observed the following order of superiority in cytoplasmic restoration and seed producibility, A2 >A1>A3 >A4. The A2 cytoplasm can therefore reliably offer seed producibility advantage to commercial seed producers. Furthermore, the which-won–where model for hybrid grain yield revealed specific adaptations of hybrid genotypes G143, G195 and G215 to first mega environment (E1 and E3), whereas G167, G110, and G112 were adapted to the second mega-environment (E2 and E4). With a GGE biplot goodness of fit of 70% the best possible yield- and stability-based ranking of hybrid genotypes was G143, G215, G189, G200, G69, G162, G119, G54, and G81. The newly uncovered hybrid products should therefore be considered for downstream testing stages and candidates for release and commercial cultivation. Clearly, this work resulted in novel findings particularly in terms of seed producibility, universal restorers across the 4 cytoplasms and environments, and superior and stable hybrid products, all of which are expected to add value to conventional and heterosis breeding in the process of sorghum cultivar development.

Assembly and comparative analysis of the mitochondrial genome in diploid potatoes.

We report the mitochondrial genome of 39 diploid potatoes and identify a candidate ORF potentially linked to cytoplasmic male sterility in potatoes. Potato (Solanum tuberosum L.) holds a critical position as the foremost non-grain food crop, playing a pivotal role in ensuring global food security. Diploid potatoes constitute a vital genetic resource pool, harboring the potential to revolutionize modern potato breeding. Nevertheless, diploid potatoes are relatively understudied, and mitochondrial DNA can provide valuable insights into key potato breeding traits such as CMS. In this study, we examine and assemble the mitochondrial genome evolution and diversity of 39 accessions of diploid potatoes using high-fidelity (HiFi) sequencing. We annotated 54 genes for all the investigated accessions, comprising 34 protein-coding genes, 3 rRNA genes, and 17 tRNA genes. Our analyses revealed differences in repeats sequences between wild and cultivated landraces. To understand the evolution of diploid maternal lineage inheritance, we conducted phylogenetic analysis, which clearly distinguished mitochondrial from nuclear gene trees, further supporting the evidence-based of clustering between wild and cultivated landraces accessions. Our study discovers new candidate ORFs associated with CMS in potatoes, including ORF137, which is homologous to other CMS in Solanaceae. Ultimately, this work bridges the gap in mitochondrial genome research for diploid potatoes, providing a steppingstone into evolutionary studies and potato breeding.

Identification and variation of a new restorer of fertility gene that induces cleavage in orf138 mRNA of Ogura male sterility in radish

Key messageA new restorer of fertility gene, Rfs, of Ogura cytoplasmic male sterility (CMS) in radish encodes a pentatricopeptide repeat protein that binds to 15 nucleotides in mRNA of the CMS gene, orf138. Nucleotide substitutions in both Rfs and orf138 determine effectiveness and specificity of restoration.Cytoplasmic male sterility (CMS) in plants caused by the expression of abnormal mitochondrial genes results from impaired pollen production. The manifestation of CMS is suppressed by the restorer of fertility (Rf) genes in the nuclear genome. Thus, the CMS-Rf system is a suitable model for studying the direct interactions of mitochondrial and nuclear genes. At least nine haplotypes, of which Type B is ancestry, have been reported for the Ogura CMS gene, orf138, in radish (Raphanus sativus). We previously observed that Rfo encoding a pentatricopeptide repeat (PPR) protein, ORF687, which inhibits the translation of orf138 is ineffective in one haplotype (i.e., Type H). Here, we carried out map-based cloning of another Rf gene (Rfs) that cleaves the orf138 mRNA of Type H. Rfs produces a PPR protein consisting of 15 PPR motifs that binds to the mRNA, cleaving the mRNA at about 50nt downstream of the binding site. However, Rfs was ineffective for Type A because of a single nucleotide substitution in the binding site. Both Rfo and Rfs suppress orf138 expression in ancestral Type B, but they are rendered ineffective in Type H and Type A, respectively, by a single nucleotide substitution in orf138.

Improvement of Quality and Disease Resistance for a Heavy-Panicle Hybrid Restorer Line, R600, in Rice (Oryza sativa L.) by Gene Pyramiding Breeding.

The utilization of heavy-panicle hybrid rice exemplifies the successful integration of architectural enhancement and heterosis, which has been widely adopted in the southwest rice-producing area of China. Iterative improvement in disease resistance and grain quality of heavy-panicle hybrid rice varieties is crucial to promote their sustainable utilization. Here, we performed a molecular design breeding strategy to introgress beneficial alleles of broad-spectrum disease resistance and grain quality into a heavy-panicle hybrid backbone restorer line Shuhui 600 (R600). We successfully developed introgression lines through marker-assisted selection to pyramid major genes (Wxb + ALKA-GC + Pigm + Xa23) derived from three parents (Huanghuazhan, I135, I488), which significantly enhance grain quality and confer resistance to rice blast and bacterial blight (BB). The improved parental R600 line (iR600) exhibited superior grain quality and elevated disease resistance while maintaining the heavy-panicle architecture and high-yield capacity of R600. Moreover, the iR600 was crossed with male sterility line 608A to obtain a new heavy-panicle hybrid rice variety with excellent eating and cooking quality (ECQ) and high yield potential. This study presents an effective breeding strategy for rice breeders to expedite the improvement of grain quality and disease resistance in heavy-panicle hybrid rice.