Male Sterility Research Articles

Restorer of fertility like 30, encoding a mitochondrion-localized pentatricopeptide repeat protein, regulates wood formation in poplar.

Nuclear-mitochondrial communication is crucial for plant growth, particularly in the context of cytoplasmic male sterility (CMS) repair mechanisms linked to mitochondrial genome mutations. The restorer of fertility-like (RFL) genes, known for their role in CMS restoration, remain largely unexplored in plant development. In this study, we focused on the evolutionary relationship of RFL family genes in poplar specifically within the dioecious Salicaceae plants. PtoRFL30 was identified to be preferentially expressed in stem vasculature, suggesting a distinct correlation with vascular cambium development. Transgenic poplar plants overexpressing PtoRFL30 exhibited a profound inhibition of vascular cambial activity and xylem development. Conversely, RNA interference-mediated knockdown of PtoRFL30 led to increased wood formation. Importantly, we revealed that PtoRFL30 plays a crucial role in maintaining mitochondrial functional homeostasis. Treatment with mitochondrial activity inhibitors delayed wood development in PtoRFL30-RNAi transgenic plants. Further investigations unveiled significant variations in auxin accumulation levels within vascular tissues of PtoRFL30-transgenic plants. Wood development anomalies resulting from PtoRFL30 overexpression and knockdown were rectified by NAA and NPA treatments, respectively. Our findings underscore the essential role of the PtoRFL30-mediated mitochondrion-auxin signaling module in wood formation, shedding light on the intricate nucleus-organelle communication during secondary vascular development.

CYTOSOLIC INVERTASE2 regulates flowering and reactive oxygen species-triggered programmed cell death in tomato.

Cytosolic invertase (CIN) in plants hydrolyzes sucrose into fructose and glucose, influencing flowering time and organ development. However, the underlying molecular mechanisms remain elusive. Through expressional, genetic, and histological analyses, we identified a substantially role of SlCIN2 (localized in mitochondria) in regulating flowering and pollen development in tomato (Solanum lycopersicum). The overexpression of SlCIN2 resulted in increased hexose accumulation and decreased sucrose and starch content. Our findings indicated that SlCIN2 interacts with Sucrose transporter2 (SlSUT2) to inhibit the sucrose transport activity of SlSUT2, thereby suppressing sucrose content in flower buds and delaying flowering. We found that higher levels of glucose in SlCIN2-overexpressing anthers result in the accumulation of abscisic acid (ABA) and reactive oxygen species (ROS), thereby disrupting programmed cell death (PCD) in anthers and delaying the end of tapetal degradation. Exogenous sucrose partially restored fertility in SlCIN2-overexpressing plants. This study revealed the mechanism by which SlCIN2 regulates pollen development and demonstrated a strategy for creating sugar-regulated gene male sterility lines for tomato hybrid seed production.

Pentatricopeptide repeat 153 (PPR153) restores maize C-type cytoplasmic male sterility in conjunction with RF4.

Maize (Zea mays L.) C-type cytoplasmic male sterility (CMS-C) is a highly used CMS system for maize commercial hybrid seed production. Rf4 is the major dominant restorer gene for CMS-C. Inbreds were recently discovered which contain the restoring Rf4 allele yet are unable to restore fertility due to the lack of an additional gene required for Rf4's restoration. To find this additional gene, QTL mapping and positional cloning were performed using an inbred that contained Rf4 but was incapable of restoring CMS-C. The QTL was mapped to a 738-kb interval on chromosome 2, which contains a Pentatricopeptide Repeat (PPR) gene cluster. Allele content comparisons of the inbreds identified three potential candidate genes responsible for fertility restoration in CMS-C. Complementation via transformation of these three candidate genes showed that PPR153 (Zm00001eb114660) is required for Rf4 to restore fertility to tassels. The PPR153 sequence is present in B73 genome, but it is not capable of restoring CMS-C without Rf4. Analysis using NAM lines revealed that Rf4 requires the presence of PPR153 to restore CMS-C in diverse germplasms. This research uncovers a major CMS-C genetic restoration pathway and can be used for identifying inbreds suitable for maize hybrid production with CMS-C cytoplasm.

GLUTAMYL-tRNA SYNTHETASE 1 deficiency confers thermosensitive male sterility in rice by affecting reactive oxygen species homeostasis.

Temperature is one of the key environmental factors influencing crop fertility and yield. Understanding how plants sense and respond to temperature changes is, therefore, crucial for improving agricultural production. In this study, we characterized a temperature-sensitive male sterile mutant in rice (Oryza sativa), glutamyl-tRNA synthetase 1-2 (ers1-2), that shows reduced fertility at high temperatures and restored fertility at low temperatures. Mutation of ERS1 resulted in severely delayed pollen development and meiotic progression at high temperatures, eventually leading to male sterility. Moreover, meiosis-specific events, including synapsis and crossover formation, were also delayed in ers1-2 compared with the wild type. However, these defects were all mitigated by growing ers1-2 at low temperatures. Transcriptome analysis and measurement of ascorbate, glutathione, and hydrogen peroxide (H2O2) contents revealed that the delayed meiotic progression and male sterility in ers1-2 were strongly associated with changes in reactive oxygen species (ROS) homeostasis. At high temperatures, ers1-2 exhibited decreased accumulation of ROS scavengers and overaccumulation of ROS. In contrast, at low temperatures, the antioxidant system of ROS was more active, and ROS contents were lower. These data suggest that ROS homeostasis in ers1-2 is disrupted at high temperatures but restored at low temperatures. We speculate that ERS1 dysfunction leads to changes in ROS homeostasis under different conditions, resulting in delayed or rescued meiotic progression and thermosensitive male fertility. ers1-2 may hold great potential as a thermosensitive material for crop heterosis breeding.

High quality RNA extraction protocol for polyphenolics-rich Cotton tissue

The extraction of high-quality RNA from cotton (Gossypium spp.) is challenging because of the presence of high polyphenolics, polysaccharides, quinones, and other secondary metabolites. A high-throughput RNA extraction protocol is a prerequisite. This Triton-X-100-based RNA extraction method utilizes Polyvinyl pyrrolidone polymer (PVPP) treatment which efficiently removes phenolics, and the application of Lithium chloride (LiCl) has been found that successfully precipitated the high-quality RNA from cotton tissue. Cytoplasmic male sterility (CMS) is a maternally inherited trait associated with specific mitochondrial genome rearrangements or mutations. The suitability of RNA extracted from Cotton CMS lines was assessed. cDNA was synthesized from RNA and assayed for mitochondrial genes (cox3, nad3, nad9) associated with male sterility. This paper discuss the advantages and limitation of this protocol over existing protocol for RNA extraction for polyphenolics-rich plant tissue.

Unbalanced lipid metabolism in anther, especially the disorder of the alpha-linolenic acid metabolism pathway, leads to cotton male sterility

Recent studies have shown that lipid metabolism is a key factor affecting anther development and male fertility. However, how plants regulating the metabolic balance of multiple lipids to ensure proper anther development and male fertility remains unclear. Analyzing lipid molecules related to anther fertility and genes responsible for their biosynthesis is crucial for understanding the physiological significance of lipid metabolism in crop fertility. In this study, we compared the transcriptome and the composition and content of lipids in anthers of two Upland cotton (Gossypium hirsutum) materials, Shida 98 (WT) and its nearly-isogenic male sterile line Shida 98A (MS). Transcriptomics analysis identified many differentially expressed genes between the two materials, with the genes of the alpha-linolenic acid metabolism pathway being the most significantly associated with the male sterility phenotype. Investigations on lipids revealed that the MS anthers over-accumulated free fatty acids (FFAs), phosphatidic acid (PA), mono- and di-galactosyldiacylglycerol (MGDG and DGDG), and had a decreased content of triacylglycerol (TAG), which was closely related to the abnormal metabolism of alpha-linolenic acid (C18:3); therefore, the major lipids containing C18:3-acyl chains, such as PA, MGDG, DGDG and TAG, are proposed to play a major role in cotton anther development. We also showed that an excessive level of MGDG and DGDG caused JA overaccumulation in MS anthers, which in turn inhibited the expression of GhFAD3 and consequently reduced the C18:3 content, presumably via a feedback regulation mechanism, ultimately affecting plant fertility. Together, our results revealed the importance of a balanced lipid metabolism in regulating the development of cotton anther and pollen and consequently male fertility.

Functional analysis of the CaPIPLC5 gene in the regulation of the fertility restoration in pepper.

Cytoplasmic male sterility (CMS) is a very important factor to produce hybrid seeds, and the restoration of fertility involves the expression of many fertility-related genes. Our previous study showed that the expression of CaPIPLC5 was significantly up-regulated in pepper restorer accessions and minimally expressed in sterile accessions, speculating that CaPIPLC5 is related to the restoration of fertility. In this study, we further validated the function of CaPIPLC5 in the restoration of fertility. The results showed that CaPIPLC5 was specifically expressed in the anthers of the restorer accessions with the subcellular localization in the cytoplasm. Furthermore, the expression of CaPIPLC5 was significantly higher in restorer lines and restorer combinations than that in CMS lines and their maintainer lines. Silencing CaPIPLC5 led to the number of pollen decreased, pollen grains wrinkled, and the ratio of pollen germination reduced. In addition, the joint analysis of Yeast One-Hybrid (Y1H) and Dual-Luciferase (dual-LUC) assays suggested that transcription factors such as CaARF5, CabZIP24 and CaMYB-like1, interacted with the promoter regions of CaPIPLC5, which regulated the expression of CaPIPLC5. The present results provide new insights into the study of CaPIPLC5 involved in the restoration of fertility in pepper.

New Observations of the Effects of the Cytoplasm of Aegilops kotschyi Boiss. in Bread Wheat Triticum aestivum L.

The cytoplasm of Aegilops kotschyi is known for the induction of male sterility and haploidy in wheat. Both systems originally appeared rather simple, but manipulation of the standard chromosome constitution of the nuclear genome revealed additional interactions. This study shows that while there is little or no allelic variation at the main fertility restorer locus Rfmulti on chromosome arm 1BS, additional genes may also be involved in the nuclear-mitochondrial genome interactions, affecting not only male fertility but also the growth rate, from pollen competition for fertilization and early endosperm divisions all the way to seed size and plant maturity. Some of these effects appear to be of a sporophytic nature; others are gametophytic. Induction of parthenogenesis by a rye inducer in conjunction with the Ae. kotschyi cytoplasm is well known. However, here we show that the cytoplasmic-nuclear interactions affect all aspects of double fertilization: producing maternal haploids from unfertilized eggs, diploids from fertilized eggs or synergids, embryo-less kernels, and fertilized eggs without fertilization of the double nucleus in the embryo sack. It is unclear how frequent the inducers of parthenogenesis are, as variation, if any, is obscured by suppressors present in the wheat genome. Genetic dissection of a single wheat accession revealed five distinct loci affecting the rate of maternal haploid production: four acting as suppressors and one as an enhancer. Only when the suppressing haplotypes are confirmed may it be possible to the identify genetic variation of haploidy inducers, map their position(s), and determine their nature and the mode of action.

Whole transcriptome analysis identifies differentially expressed mRNA, miRNA and lncRNA associated with male sterility in the silkworm, Bombyx mori

Insect sterility technology is gradually being applied to the control of lepidoptera pests, and the target gene for male sterility is the core of this technology. JMS is a mutant silkworm that exhibits male sterility, and to elucidate its formation mechanism, this study conducted a full transcriptome analysis of the testes of JMS and its wild-type silkworms 48 h after pupation, identifying 205 DElncRNAs, 913 mRNAs, and 92 DEmiRNAs. The KEGG pathway enrichment analysis of the DEmRNAs revealed that they were involved in the biosynthesis of amino acids and ECM-receptor interactions. Combined with ceRNA regulatory network KEGG analysis suggests that pathways from amino acid biosynthesis to hydrolytic processes of protein synthesis may play a crucial role in the formation of JMS mutant variants. Our study deepens our understanding of the regulatory network of male sterility genes in silkworms; it also provides a new perspective for insect sterility technology.

Marker Assisted Characterization of TGMS Gene in Rice Cultigen Ts 29 (Oryza Sativa L.) to Develop Two Line Hybrids

The application of the thermsensitive genic male sterility (TGMS) system has great potential for revolutionizing hybrid rice production through simple and effective seed production technology. For the successful utilization of this novel male sterility system, knowledge of the fertility behaviour and genetic control of a TGMS line is essential. In this study, an investigation was carried out to characterize the TGMS gene present in the mapping population of TS 29 x CO(R)49. Through Bulk Segregant Analysis RM 3476 alone produced a polymorphic relationship between the parents and corresponding bulks and was used for surveying the individual plants of F3/F4 population. The marker RM 3476 produced contrasting fragments between sterile and fertile plants. The result was associated with the phenotypic observations on pollen fertility under the field conditions, confirming co-segregation of the marker with the gene of interest. Hence, the tight linkage of marker with TGMS gene was identified by RM 3476 which was already identified as a marker located near tms 6 gene in chromosome 5. From this study four TGMS lines were also identified with desirable floral traits for high out crossing and medium slender grain type. Bangladesh J. Bot. 53(2): 345-351, 2024 (June)

Controlled Environment Ecosystem: A Cutting-Edge Technology in Speed Breeding.

The controlled environment ecosystem is a meticulously designed plant growing chamber utilized for cultivating biofortified crops and microgreens, addressing hidden hunger and malnutrition prevalent in the growing population. The integration of speed breeding within such controlled environments effectively eradicates morphological disruptions encountered in traditional breeding methods such as inbreeding depression, male sterility, self-incompatibility, embryo abortion, and other unsuccessful attempts. In contrast to the unpredictable climate conditions that often prolong breeding cycles to 10-15 years in traditional breeding and 4-5 years in transgenic breeding within open ecosystems, speed breeding techniques expedite the achievement of breeding objectives and F1-F6 generations within 2-3 years under controlled growing conditions. In comparison, traditional breeding may take 5-10 years for plant population line creation, 3-5 years for field trials, and 1-2 years for variety release. The effectiveness of speed breeding in trait improvement and population line development varies across different crops, requiring approximately 4 generations in rice and groundnut, 5 generations in soybean, pea, and oat, 6 generations in sorghum, Amaranthus sp., and subterranean clover, 6-7 generations in bread wheat, durum wheat, and chickpea, 7 generations in broad bean, 8 generations in lentil, and 10 generations in Arabidopsis thaliana annually within controlled environment ecosystems. Artificial intelligence leverages neural networks and algorithm models to screen phenotypic traits and assess their role in diverse crop species. Moreover, in controlled environment systems, mechanistic models combined with machine learning effectively regulate stable nutrient use efficiency, water use efficiency, photosynthetic assimilation product, metabolic use efficiency, climatic factors, greenhouse gas emissions, carbon sequestration, and carbon footprints. However, any negligence, even minor, in maintaining optimal photoperiodism, temperature, humidity, and controlling pests or diseases can lead to the deterioration of crop trials and speed breeding techniques within the controlled environment system. Further comparative studies are imperative to comprehend and justify the efficacy of climate management techniques in controlled environment ecosystems compared to natural environments, with or without soil.

Eliminating malaria vectors with precision-guided sterile males

Controlling the principal African malaria vector, the mosquito Anopheles gambiae, is considered essential to curtail malaria transmission. However, existing vector control technologies rely on insecticides, which are becoming increasingly ineffective. Sterile insect technique (SIT) is a powerful suppression approach that has successfully eradicated a number of insect pests, yet the A. gambiae toolkit lacks the requisite technologies for its implementation. SIT relies on iterative mass releases of nonbiting, nondriving, sterile males which seek out and mate with monandrous wild females. Once mated, females are permanently sterilized due to mating-induced refractoriness, which results in population suppression of the subsequent generation. However, sterilization by traditional methods renders males unfit, making the creation of precise genetic sterilization methods imperative. Here, we introduce a vector control technology termed precision-guided sterile insect technique (pgSIT), in A. gambiae for inducible, programmed male sterilization and female elimination for wide-scale use in SIT campaigns. Using a binary CRISPR strategy, we cross separate engineered Cas9 and gRNA strains to disrupt male-fertility and female-essential genes, yielding >99.5% male sterility and >99.9% female lethality in hybrid progeny. We demonstrate that these genetically sterilized males have good longevity, are able to induce sustained population suppression in cage trials, and are predicted to eliminate wild A. gambiae populations using mathematical models, making them ideal candidates for release. This work provides a valuable addition to the malaria genetic biocontrol toolkit, enabling scalable SIT-like confinable, species-specific, and safe suppression in the species.

Isolation and evaluation of novel male sterile and self‐incompatible mutant lines of mungbean (Vigna radiata L.)

AbstractHeterosis has long been harnessed in crop production, and while crop fertility lays the groundwork for leveraging heterosis, there remains a dearth of comprehensive data regarding genic male sterility in mungbean. Mungbean (Vigna radiata (L.) Wilczek), being a self‐pollinating crop, exhibits significant hybrid vigour in F1 hybrid seed yield, suggesting the potential for developing hybrid varieties to propel mungbean yield beyond existing plateaus. A new male sterile mutant was induced, isolated, stabilized and evaluated, via gamma irradiation at a rate of 600 Gy, identified in the M3 generation of the GM 4 accession. A uniform male sterile line was crossbred with the wild‐type parental plant to explore the inheritance pattern of male sterility. Pollen sterility was validated through the acetocarmine test, while stigma viability was ascertained using the hydrogen peroxide. While all F1 plants displayed fertile flowers, the F2 generation showed a clear 3:1 segregation ratio for fertile to male sterile plants, indicating the control of male sterility by a single recessive gene, mms. Also, a novel existence of a truncated stigma nestled within the anther column inhibits the effective reception of pollen during anthesis, offering potential in reducing emasculation time by a slight cut on the flower bud and enhancing cross pollination during hybrid seed production. The isolation of these two mutants is poised to significantly advance the global mungbean hybrid breeding programme.

Exploration of the potential of a boosted sterile insect technique to control fruit flies in mango orchards.

An innovative version of the sterile insect technique (SIT) for pest control, called boosted SIT, relies on the use of sterile males coated with a biocide to control a target wild pest population of the same species. The objective of the present study was to assess the relevance of such technology to control the fruit fly Bactrocera dorsalis and fruit losses in mango orchards using. An agent-based simulation model named BOOSTIT was used to explore the reduction of fruit losses thank to sterile male fruit flies control and economic benefits according to different strategies of sterile male release. The simulation considered a landscape of 30.25 ha made up of four mango orchards. The SIT and the boosted SIT reduced fruit losses when releases were made before the mango fruiting period. According to model simulations, releases should be performed at least seven times at 2-week intervals and with a sterile/wild male ratio of at least 10:1. Considering the benefit/cost ratio (BCR), few releases should be done with a late start date. The BCR showed economic gains from the two control methods, the number of saved fruits and BCR being higher for SIT. Our simulations showed that SIT would have better results than the boosted SIT to contribute to an effective control of Bactrocera dorsalis at the scale of a small landscape. We highlight the need for laboratory studies of other types of pathogen to find a suitable one with higher incubation time and lower cost. © 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Mark-Release-Recapture (MRR) of Sterile Male Aedes albopictus (Skuse) in Sri Lanka: Field Performance of Sterile Males and Estimation of the Wild Mosquito Population Density.

Dengue is an important mosquito-borne disease in Sri Lanka. The Sterile Insect Technique (SIT) is an environment-friendly and novel method that can suppress dengue vector mosquitoes in Sri Lanka. This study aimed to evaluate the field performance of sterile males and the density of wild male Aedes albopictus (Skuse) using a Mark-Release-Recapture (MRR) assay. Laboratory-colonized male pupae were exposed to 50 Gy gamma using a Co60 source. Sterile males (approx. 10,000) marked with fluorescent dust were released weekly for 4 consecutive weeks (January-February 2021) in a geographically isolated 30 ha site in Gampaha. Results show sterile males could disperse up to 543.8 m with a mean distance of 255.1 ± 44.6 m and survive up to 6 days with a mean life expectancy of 3.55 ± 2.32 days. A high field mating competitiveness of sterile males based on a Fried value of 0.47 ± 0.007 and significant induced sterility in the wild eggs in the second generation were found. The mean wild male mosquito population density was 163 males/ha. The data generated will be useful for designing future trials in Sri Lanka and other countries with similar situations.

TENT5-mediated polyadenylation of mRNAs encoding secreted proteins is essential for gametogenesis in mice

Cytoplasmic polyadenylation plays a vital role in gametogenesis; however, the participating enzymes and substrates in mammals remain unclear. Using knockout and knock-in mouse models, we describe the essential role of four TENT5 poly(A) polymerases in mouse fertility and gametogenesis. TENT5B and TENT5C play crucial yet redundant roles in oogenesis, with the double knockout of both genes leading to oocyte degeneration. Additionally, TENT5B-GFP knock-in females display a gain-of-function infertility effect, with multiple chromosomal aberrations in ovulated oocytes. TENT5C and TENT5D both regulate different stages of spermatogenesis, as shown by the sterility in males following the knockout of either gene. Finally, Tent5a knockout substantially lowers fertility, although the underlying mechanism is not directly related to gametogenesis. Through direct RNA sequencing, we discovered that TENT5s polyadenylate mRNAs encoding endoplasmic reticulum-targeted proteins essential for gametogenesis. Sequence motif analysis and reporter mRNA assays reveal that the presence of an endoplasmic reticulum-leader sequence represents the primary determinant of TENT5-mediated regulation.

Gibberellin Signaling through RGA Suppresses GCN5 Effects on Arabidopsis Developmental Stages.

Histone acetyltransferases (HATs) modify the amino-terminal tails of the core histone proteins via acetylation, regulating chromatin structure and transcription. GENERAL CONTROL NON-DEREPRESSIBLE 5 (GCN5) is a HAT that specifically acetylates H3K14 residues. GCN5 has been associated with cell division and differentiation, meristem function, root, stem, foliar, and floral development, and plant environmental response. The flowers of gcn5 plants display a reduced stamen length and exhibit male sterility relative to the wild-type plants. We show that these effects may arise from gibberellin (GA)-signaling defects. The signaling pathway of bioactive GAs depends on the proteolysis of their repressors, DELLA proteins. The repressor GA (RGA) DELLA protein represses plant growth, inflorescence, and flower and seed development. Our molecular data indicate that GCN5 is required for the activation and H3K14 acetylation of genes involved in the late stages of GA biosynthesis and catabolism. We studied the genetic interaction of the RGA and GCN5; the RGA can partially suppress GCN5 action during the whole plant life cycle. The reduced elongation of the stamen filament of gcn5-6 mutants is reversed in the rga-t2;gcn5-6 double mutants. RGAs suppress the GCN5 effect on the gene expression and histone acetylation of GA catabolism and GA signaling. Interestingly, the RGA and RGL2 do not suppress ADA2b function, suggesting that ADA2b acts downstream of GA signaling and is distinct from GCN5 activity. In conclusion, we propose that the action of GCN5 on stamen elongation is partially mediated by RGA and GA signaling.

Identification, Elucidation and Deployment of a Cytoplasmic Male Sterility System for Hybrid Potato.

Recent advances in diploid F1 hybrid potato breeding rely on the production of inbred lines using the S-locus inhibitor (Sli) gene. As a result of this method, female parent lines are self-fertile and require emasculation before hybrid seed production. The resulting F1 hybrids are self-fertile as well and produce many undesirable berries in the field. Utilization of cytoplasmic male sterility would eliminate the need for emasculation, resulting in more efficient hybrid seed production and male sterile F1 hybrids. We observed plants that completely lacked anthers in an F2 population derived from an interspecific cross between diploid S. tuberosum and S. microdontum. We studied the antherless trait to determine its suitability for use in hybrid potato breeding. We mapped the causal locus to the short arm of Chromosome 6, developed KASP markers for the antherless (al) locus and introduced it into lines with T and A cytoplasm. We found that antherless type male sterility is not expressed in T and A cytoplasm, proving that it is a form of CMS. We hybridized male sterile al/al plants with P cytoplasm with pollen from al/al plants with T and A cytoplasm and we show that the resulting hybrids set significantly fewer berries in the field. Here, we show that the antherless CMS system can be readily deployed in diploid F1 hybrid potato breeding to improve hybridization efficiency and reduce berry set in the field.

CWF19L2 is Essential for Male Fertility and Spermatogenesis by Regulating Alternative Splicing.

The progression of spermatogenesis along specific developmental trajectories depends on the coordinated regulation of pre-mRNA alternative splicing (AS) at the post-transcriptional level. However, the fundamental mechanism of AS in spermatogenesis remains to be investigated. Here, it is demonstrated that CWF19L2 plays a pivotal role in spermatogenesis and male fertility. In germline conditional Cwf19l2 knockout mice exhibiting male sterility, impaired spermatogenesis characterized by increased apoptosis and decreased differentiated spermatogonia and spermatocytes is observed. That CWF19L2 interacted with several spliceosome proteins to participate in the proper assembly and stability of the spliceosome is discovered. By integrating RNA-seq and LACE-seq data, it is further confirmed CWF19L2 directly bound and regulated the splicing of genes related to spermatogenesis (Znhit1, Btrc, and Fbxw7) and RNA splicing (Rbfox1, Celf1, and Rbm10). Additionally, CWF19L2 can indirectly amplify its effect on splicing regulation through modulating RBFOX1. Collectively, this research establishes that CWF19L2 orchestrates a splicing factor network to ensure accurate pre-mRNA splicing during the early steps of spermatogenesis.

Trait Correlation and Path Study for Quantitative Traits in Garlic (Allium sativum L.) Genotypes

Garlic is a widely cultivated vegetable with both culinary and medicinal uses in Ethiopia. The analyses, focusing on various traits, help in identifying genotypes with high productivity and quality. Garlic reproduces solely vegetatively due to male and female sterility in its gametophytes. Its domestication originated from the wild species Allium longicuspis Regel. The introduction of diverse garlic varieties contributes to enhancing its genetic potential. This study aimed to assess the relationship between different traits and their influence on garlic bulb yield. The experiment conducted during the rainy season of 2020–2021 at the Fogera National Rice Research and Training Center in Ethiopia. The experiment included 49 garlic genotypes arranged in a simple lattice design with two replications. The results revealed significant genotypic correlation coefficients between fresh bulb yield per hectare and bulb weight per plant (r = 0.82***), pseudo-stem height (r = 0.82***), and clove weight (r = 0.81***). Similarly, significant positive phenotypic associations were observed between fresh bulb yield per hectare and clove weight per bulb (r = 0.77***), pseudo-stem height (r = 0.77***), and bulb weight per plant (r = 0.76***). Further analysis indicated that pseudo-stem height (0.42) and clove weight (0.39) exerted the highest phenotypic direct effects on fresh bulb yield per hectare. Therefore, traits such as clove weight, pseudo-stem height, and bulb weight exhibit high genotypic coefficients and significant direct effects on total bulb yield, making them crucial for direct selection to increase bulb yield.