F1 Progeny Research Articles

Selection, inheritance, life-history traits, and flight capacity of Grapholita molesta (Lepidoptera: Tortricidae) resistant to lambda-cyhalothrin.

Grapholita molesta (Busck) (Lepidoptera: Tortricidae) is a globally significant fruit pest traditionally managed using chemical insecticides such as lambda-cyhalothrin. However, repeated applications have led to the development of resistance in field populations. The mechanisms and fitness costs associated with lambda-cyhalothrin resistance in G. molesta remain poorly understood. In this study, we established a lambda-cyhalothrin-resistant strain (RS) of G. molesta through continuous selection with lambda-cyhalothrin in the laboratory for 21 generations, resulting in a 249.6-fold increase in resistance. Bioassay data from the F1 progeny (F1RS and F1SR) of reciprocal crosses between RS and susceptible strain (SS) showed similar degrees of dominance (0.40 for F1RS and 0.32 for F1SR). Significant differences between observed and expected mortalities of backcross individuals suggested that resistance to lambda-cyhalothrin is polygenic. Compared to the SS strain, the RS strain exhibited significantly prolonged periods of larval, prepupal, and pupal stages, as well as a significant decrease in larval weight. In addition, the RS strain showed significant reductions in fecundity, cumulative flight distance, maximum flight distance, cumulative flight time, maximum flight time, and average flight speed. These results suggest that resistance to lambda-cyhalothrin in G. molesta is autosomal, incompletely dominant, and polygenic. Life-history traits such as fecundity and flight capacity were significantly reduced in RS strains compared with the SS strain. These findings provide valuable insights for mitigating lambda-cyhalothrin resistance development and promoting sustainable control of G. molesta in the field.

Transgenerational Effects on Lifespan and Pathology of Paternal Pre-conceptional Exposure to Continuous Low-dose-rate Gamma Rays in C57BL/6J Mice

The present work investigates the multigenerational effects of paternal pre-conceptional exposure to continuous low-dose-rate gamma rays in C56BL/6J mice. Male C57BL/6J (F0 sires) mice were exposed to low dose rates of 20, 1, and 0.05 mGy/day for 400 days, to total accumulated doses of 8,000, 400, and 20 mGy, respectively. Upon completion of the radiation exposure, the F0 male mice were immediately bred to non-irradiated 8-week-old C57BL/6J females (F0 dams) to produce the first-generation (F1) mice. Randomly selected F1 males and females were then bred to produce the second-generation (F2) mice. All the mice, except the F0 dams, were subjected to pathological examination upon natural death. Reproductive parameters, lifespan, causes of death, neoplasm incidences and non-neoplastic disease incidences were used as parameters to evaluate the biological effects of continuous pre-conceptional exposure of the sires (F0) to continuous low-dose-rate radiation. There were no significant differences in the pregnancy and weaning rates among the parent (F0) generation. Average litter size and average number of weaned pups (F1) from dams bred to males (F0) exposed to 20 mGy/day were significantly decreased compared to the non-irradiated controls. Significant lifespan shortening in the sires (F0) was observed only in the 20 mGy/day group due to early death from malignant lymphomas. Life shortening was also observed in the F1 progeny of sires (F0) exposed to 20 and 1 mGy/day, but could not be attributed to a specific cause. No significant differences in the causes of death were found between dose groups in any generation. The number of primary tumors per mouse was significantly increased only in the F0 males exposed to 20 mGy/day. Except for the increased incidence rate for Harderian gland neoplasms in sires (F0) exposed to 20 mGy/day, there was no significant difference in neoplasm incidences and tumor spectra in all 3 generations in each sex regardless of radiation exposure. No multi- or transgenerational effects in the parameters examined were observed in the F1 and F2 progeny of sires exposed to 0.05 mGy/day for 400 days.

Toxicity and Sublethal Effect of Chlorantraniliprole on Multiple Generations of Aedes aegypti L. (Diptera: Culicidae)

Due to the quick development of insecticide resistance, it is crucial to optimize management programs by understanding the sublethal effects of effective insecticides like chlorantraniliprole on Aedes aegypti L. populations. Using age-stage and two-sex life tables, we investigated the sublethal impacts of chlorantraniliprole on Ae. aegypti. Larval duration in the progeny of exposed parents was reduced by 0.33–0.42 days, whereas, the longevity of male and female adults was decreased by 1.43–3.05 days. Similarly, the egg-laying capacity of F1 and F2 progeny of the exposed parents was significantly reduced from 27.3% to 41.2%. The mean generation time (T) increased up to 11.8% in exposed populations, and the net reproduction rate (Ro) decreased by 51.50–55.60%. After 24 h of chlorantraniliprole treatment, there was a significant increase in cytochrome P450 activity. Contrarily, the activity of glutathione S-transferase (GST) initially declined but started increasing after 48 h of treatment. This research highlights the importance of chlorantraniliprole in mosquito management, as well as the importance of considering sublethal effects when developing strategies to handle them. Having a thorough understanding of the harmful effects of insecticides on mosquito populations can greatly enhance the effectiveness of insecticide-based interventions, while also minimizing the risk of pest resurgence.

Combining Ability of Capsicum annuum Hybrid for Antioxidant Activities, Polyphenol Content, α-Glucosidase Inhibitory, Yield, and Yield Components.

Chili (Capsicum annuum) consumption is often suggested, and using functional food cultivars is the most effective strategy post COVID-19 pandemic. Controlling chili breeding activity is one of the most effective methods to produce new hybrid varieties. However, the general combining ability (GCA), specific combining ability (SCA), and heterotic effect of functional biochemicals (polyphenol content, antioxidant activities, and α-glucosidase inhibitory compounds) remain poorly known in C. annuum. This study aimed to estimate these parameters in C. annuum by using five different genotypes and their hybrid combinations based on growth characteristics, yield, yield components, and fruit functional biochemicals. The F1 and F1R progenies were obtained from crosses in a greenhouse with a full diallel mating design. Each parent used in this study had a GCA advantage for each characteristic. The hybrid combination of IPB074 × IPB005 and IPB435 × IPB367 displayed the best yield results. However, the results indicated the opposite regarding α-glucosidase inhibitory compounds. The heterotic effect of functional biochemicals was observed for traits related to genotypes, polyphenol content, antioxidant activity, α-glucosidase inhibitory compounds, and similar properties related to yield and yield components, indicating their use in hybrid chili production.

Susceptibility of olive cultivars and selections to Colletotrichum species causing anthracnose in Spain.

Anthracnose, the most critical fruit disease affecting olive crops, necessitates the evaluation of the susceptibility of traditional and new varieties. In Spain and Portugal, Anthracnose is caused by several Colletotrichum species, with C. godetiae and C. nymphaeae being dominant and C. acutatum and C. fioriniae being secondary. This study explores the susceptibility of fruits from an F1 progeny resulting from a cross between 'Picual' (resistant) and 'Arbequina' (moderately susceptible) cultivars to C. godetiae. While most genotypes showed resistance levels comparable to their parents, seven showed a 50% reduction in disease severity compared to 'Picual.' The normal distribution of genotypes' response to the pathogen suggests a complex resistance mechanism. Furthermore, we assessed the susceptibility of four traditional cultivars, two new cultivars ('Sikitita-2' and 'Martina'), and five advanced selections (pre-commercial genotypes) to C. godetiae and C. nymphaeae. Despite the significant interaction between the olive genotype and Colletotrichum species in this experiment, the new cultivars and advanced selections were classified as susceptible or moderately susceptible against both species. A subsequent analysis of the interaction between 'Picual' (resistant) and 'Hojiblanca' (susceptible) fruits with the four mentioned Colletotrichum species revealed significant differences among cultivars but no interaction between genotype and pathogen species. Colletotrichum species were categorized as follows: i) C. godetiae and C. nymphaeae as highly virulent, ii) C. acutatum as moderately virulent, and iii) C. fioriniae as weakly virulent. Finally, C. nymphaeae exhibited an enhanced ability to infect and develop acervuli in olive leaves, potentially serving as an inoculum source for this species. The absence of a correlation between leaf and fruit susceptibilities to the pathogen suggests differences in resistance mechanisms. In conclusion, this study provides valuable insights into the complex interactions between olive genotype and Colletotrichum species, essential for cultivar selection and understanding the disease cycle.

Genetic Analysis for Seed Yield and Yield‐Related Traits in Tepary Bean (Phaseolus acutifolius A. Gray) Under Drought‐Stress and Non‐stress Conditions

ABSTRACTTepary bean (Phaseolus acutifolius A. Gray) is an under‐utilized genetic resource with significant potential for food security and stress tolerance breeding. Expanding its cultivation in southern Africa requires high‐yielding, locally adapted and drought‐tolerant varieties. This study determined the combining ability and genetic components for seed yield and related traits in tepary bean genotypes under non‐stress (NS) and drought‐stress (DS) conditions. Seven parents were selected through rigorous phenotyping and crossed using a half‐diallel design. The 7 parents and 21 F2 progenies were evaluated in 2021/2022 season at Kasinthula and Bunda sites in Malawi under NS and DS conditions using a 4 × 7 lattice design with three replications. The specific combining ability (SCA) × location interaction effect was significant (p < 0.05) for DTF, NPP, and SY, suggesting that the genetic effects of crosses were influenced by the test locations. General combining ability (GCA) and SCA mean squares were significant for the number of seeds per pod (NSP) and SY under DS conditions, indicating both additive and non‐additive gene effects. Baker's ratio (BR) > 0.50 for NPP and NSP under DS conditions suggested a preponderance of additive gene effects. G40145, G40148 and G40150 parental lines were good combiners for NPP and SY. The F2 families from crosses such as Zimbabwe landrace/G40138, Zimbabwe landrace/G40150, G40059/G40145, G40059/G40148, G40138/G40150 and G40145/G40150 were identified as best specific combiners, with enhanced SY of 1.67 t/ha under DS conditions. The study recommends advancing high‐performing early‐generation families for selection across representative environments to facilitate variety release and commercialization.

Marker assisted selection and genome recovery for heat tolerance in segregants of rice (Oryza sativa L.)

Abstract In the present global warming scenario, it is urgent to impart heat tolerance into the popular high yielding rice (Oryza sativa L.) varieties to reduce yield loss to a great extent. Uma (a prevalent red-grained rice variety in Kerala) was crossed with Nagina 22 (N22), the popular donor for heat tolerance. From the seven promising heat-tolerant F3 plants previously identified through marker assisted selection and field screening, F4 lines (58 nos.) were raised and characterised morphologically. Heat tolerance was scored under natural heat stress during hot summer months in field conditions. Among the evaluated F4 progenies, 26 promising plants registered >75% spikelet fertility. Double panicles were observed in some of the tillers of fourteen F4 plants. During the selection with reported polymorphic markers between parents, all the 26 F4 plants produced the corresponding amplicon for RM5749 (linked marker for heat tolerance in the parental cross Uma x N22) and also for RM9, RM201, RM208, RM225, RM242, RM495, RM3586, RM6100, RM6836, and RM26212, corresponding to the heat tolerant parent N22. Genomic analysis revealed 64% recovery of the Uma genome in seven F4 plants, with maximum genomic regions of Uma on the chromosomes 3 and 5. These promising F4 plants could be forwarded to generate stable and high yielding heat tolerant rice varieties. The identified markers could be used for linkage analysis and QTL mapping in future. Keywords: Heat stress; Rice; spikelet fertility; SSR; QTL

Embryonal exposure to 4-methylbenzylidene camphor induces reproduction impairment in adult zebrafish (Danio rerio)

This study investigated how early exposure to xenobiotics can lead to disease in adulthood, which is challenging for toxicologists. We employed a ‘cradle to grave’ approach using zebrafish (Danio rerio) embryos exposed to 4-methylbenzylidene camphor (4-MBC), a commonly used organic UV filter. Molecular docking and simulation studies confirmed the predictive toxicity and stable interaction of 4-MBC with androgen and estrogen receptors, with binding energies of −9.28 and −9.01 kcal/mol, respectively. Exposure to 4-MBC at 5, 50, and 500 μg/L concentrations resulted in significantly altered transcriptional and translational responses of ar, esr1, and vtg1 genes in embryos at 120 h post-fertilization (hpf). The exposure induced a non-monotonic dose-response pattern (NMDR), a characteristic feature of endocrine-disrupting chemicals. Additionally, a significant decrease in fertilization was observed in adults. Although fecundity was not affected in inter- and intra-breeding performances, developmental deformities were observed in F1 progenies with impaired survival at 10 days post-fertilization. The findings of this study show that embryonic exposure to 4-MBC is likely to induce reproductive and transgenerational toxicity in D. rerio and exhibit endocrine disruption in aquatic non-target organisms. This work is the first to elucidate the low-level long-term effects of 4-MBC from the embryonic stage to adulthood.

Genetic Evidence of Recessive Epistasis Among Sex Determining silkless1 (sk1) tasselseed1 (ts1) and tasselseed2 (ts2) Genes and Its Utility in Maize (Zea mays L.) Breeding

ABSTRACTIn maize, sex is determined by the formation of staminate and pistillate florets from an initially bisexual floral meristem through action of Silkless1 (Sk1), Tasselseed1 (Ts1) and Tasselseed2 (Ts2) genes. In this study, separate temperate donors having mutant sk1 (EC904829 and EC904831), ts1 (EC904827) and ts2 (EC904824) genes were crossed with sub‐tropical inbreds (HKI323 and HKI1105) to investigate the genetics and interaction of silkless and tasselseed traits. The monogenic and nuclear inheritance of sk1, ts1 and ts2 genes was established in a standard Mendelian principles in both monohybrid and dihybrid crosses irrespective of genetic background. About 444 F2 progenies across digenic population ‐A, ‐B and ‐C segregated as 239 normal: 109 silkless: 96 tasselseed, and 377 progenies of digenic populations ‐D, ‐E and ‐F segregated with 214 normal: 79 silkless: 84 tasselseed plants revealing recessive epistatic interaction (9:3:4) of sk1 with two ts mutations (ts1 and ts2). No progeny with both silkless and tasselseed phenotype was observed. The double mutants (sk1sk1/ts1ts1 and sk1sk1/ts2ts2) were identified through progeny testing. Our study emphasize the untapped potential of these mutants, providing a renewed focus for maize breeders worldwide.

Characterization of novel wheat-Thinopyrum ponticum introgression lines derived from partial amphiploid AUS6770 for resistance to stripe rust

The wild decaploid species Thinopyrum ponticum (Podp.) Barkworth & D.R. Dewey is an important source of genes against biotic and abiotic stresses affecting wheat. The wheat–Th. ponticum partial amphiploid AUS6770 shows resistance to multiple diseases, including stripe rust, stem rust, and powdery mildew. Mitotic chromosomes of AUS6770 were characterized by non-denaturing-fluorescence in situ hybridization (ND-FISH), and the individual Th. ponticum chromosomes 1Ae to 7Ae were karyotypically distinguished by Oligo-FISH painting using bulked oligo pools based on wheat-barley collinear regions. A novel stripe rust resistant line A155, derived from AUS6770, was found to have 44 chromosomes, including a pair of 2Ae chromosomes and a pair of 6B-6Ae translocations. To detect plants with transfer of resistance genes from A155 to wheat chromosomes, 1770 plants were developed from F2–F5 progenies of A155 crossed with the susceptible wheat cultivar MY11 and characterized with ND-FISH using multiple probes. A high frequency of transmission of chromosome 2Ae was observed, and 31 types of 2Ae chromosomal aberrations were identified using ND-FISH. Ten chromosomal bins on the 2Ae chromosome were determined from the deletion and translocation lines based on genome-based PCR markers. In combination with the evaluation of disease resistance, the gene(s) for stripe rust resistance was located on the FL0.79–1.00 of 2AeS and covers the corresponding region of 0–58.26 Mb in the reference genome of Th. elongatum. The newly identified wheat-Th. ponticum 2Ae translocation lines can be exploited as potential germplasm in wheat breeding for stripe rust resistance

Grain Appearance Quality of Parental and \(F_2\) Segregating Populations of Aus Rice (Oryza sativa L.)

Longer grain length with smaller grain breadth, length-breadth ratio and 1000-grain weight is considered desirable character for the development of fine grain premium quality rice. In the present experiment, grain appearance quality of 15 F2 families and 10 parental genotypes were determined to select fine grain rice. The maximum mean value for thousand seed weight was found from the genotypes P2 (29.32g), P1 (28.89g), and F2 generations of P2 x P5 (32.77g), P2 x P1 (27.30g), P1 x P4 (26.04g) and were considered as high yielders in terms of grain weight. The highest mean values for grain length were found from the genotypes P6 (9.95 mm; grain breadth 2.23 mm), P7 (9.36 mm; grain breadth 2.62 mm), and F2 generations of P6 x P7 (9.46 mm; grain breadth 2.44 mm), P3 x P6 (9.40 mm; grain breadth 2.18 mm), P7 x P6 (9.37 mm; grain breadth 2.29 mm), P7 x P3 (9.36 mm; grain breadth 2.33 mm). The F2 progenies of the cross P1 x P4 (7.79 to 11.61 mm) and P6 x P3 (7.69 to 11.11 mm) showed extremes performance for grain length indicating transgressive segregation for this trait. The superior genotypes with desirable grain characters could be selected from transgressive segregation of F2 families for the development of new varieties of fine grain rice.

Dysregulated miR-124 mediates impaired social memory behavior caused by paternal early social isolation.

Early social isolation (SI) leads to various abnormalities in emotion and behavior during adulthood. However, the negative impact of SI on offspring remains unclear. This study has discovered that paternal early SI causes social memory deficits and anxiety-like behavior in F1 young adult mice, with alterations of myelin and synapses in the medial prefrontal cortex (mPFC). The 2-week SI in the F1 progeny exacerbates social memory impairment and hypomyelination in the mPFC. Furthermore, the down-regulation of miR-124, a key inhibitor of myelinogenesis, or over-expression of its target gene Nr4a1 in the mPFC of the F1 mice improves social interaction ability and enhances oligodendrocyte maturation and myelin formation. Mechanistically, elevated levels of miR-124 in the sperm of paternal SI mice are transmitted epigenetically to offspring, altering the expression levels of miR-124/Nr4a1/glucocorticoid receptors in mPFC oligodendrocytes. This, in turn, impedes the establishment of myelinogenesis-dependent social behavior. This study unveils a novel mechanism through which miR-124 mediates the intergenerational effects of early isolation stress, ultimately impairing the establishment of social behavior and neurodevelopment.

Genetic Mapping and Characterization of the Clubroot Resistance Gene BraPb8.3 in Brassica rapa.

Clubroot, a significant soil-borne disease, severely impacts the productivity of cruciferous crops. The identification and development of clubroot resistance (CR) genes are crucial for mitigating this disease. This study investigated the genetic inheritance of clubroot resistance within an F2 progeny derived from the cross of a resistant parent, designated "377", and a susceptible parent, designated "12A". Notably, "377" exhibited robust resistance to the "KEL-23" strain of Plasmodiophora brassicae, the causative agent of clubroot. Genetic analyses suggested that the observed resistance is controlled by a single dominant gene. Through Bulked Segregant Analysis sequencing (BSA-seq) and preliminary gene mapping, we localized the CR gene locus, designated as BraPb8.3, to a 1.30 Mb genomic segment on chromosome A08, flanked by the markers "333" and "sau332-1". Further fine mapping precisely narrowed down the position of BraPb8.3 to a 173.8 kb region between the markers "srt8-65" and "srt8-25", where we identified 22 genes, including Bra020861 with a TIR-NBS-LRR domain and Bra020876 with an LRR domain. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) analyses confirmed that both Bra020861 and Bra020876 exhibit increased expression levels in the resistant parent "377" following inoculation with P. brassicae, thereby underscoring their potential as key genes implicated in BraPb8.3-mediated clubroot resistance. This study not only identifies molecular markers associated with BraPb8.3 but also enriches the genetic resources available for breeding programs aimed at enhancing resistance to clubroot.

Inheritance Analysis for Quantitative Traits in F2 Populations of Mustard Genotypes Grown Under Heat Stress Environments

Genetic variability is the basic requirement for improving crops which further leads to progress in crop breeding. This study was carried out to determine the inheritance pattern of various agronomically important traits in F2 populations of mustard genotypes. The objective of this study was to determine the genetic variability for agro-morphological traits under control conditions (CC) and heat stress environments (HS) and provide the future strategies to improve this important oilseed crop. The experiment was conducted at the Botanical Garden, Sindh Agriculture University, Tandojam. During 2022-23, a total of 27 genotypes including nine parental mustard genotypes and eighteen F2 progenies were grown under control and heat stress conditions. The experiment was laid out in a randomized complete block design with three replications. The analysis of variance for the genotypes, treatments, and F2 progenies revealed significant differences (P≤0.05) in most of the studied characters, highlighting the importance of genetic variations in the evaluated breeding materials of mustard for stress breeding. When estimating heritability for different characters, including seed yield plant-1, it was found that F2 crosses showed varying levels of heritability for different characteristics, ranging from low to moderate and high. This implies that careful selection may be conducted for traits that have low heritability, especially for seed yield.; however, those characters which showed high heritability would be selected at early generation stage to improve the characters.

Multi-Population Analysis for Leaf and Neck Blast Reveals Novel Source of Neck Blast Resistance in Rice.

Rice blast is one of the most devastating biotic stresses that limits rice productivity. The North Eastern Hill (NEH) region of India is considered to be one of the primary centres of diversity for both rice and pathotypes of Magnaporthe grisea. Therefore, the present study was carried out to elucidate the genetic basis of leaf and neck blast resistance under Meghalaya conditions. A set of 80 diverse genotypes (natural population) and 2 F2 populations involving resistant parent, a wildtype landrace, LR 5 (Lal Jangali) and susceptible genotypes Sambha Mahsuri SUB 1 (SMS) and LR 26 (Chakhao Poireiton) were used for association analysis of reported major gene-linked markers with leaf and neck blast resistance to identify major effective genes under local conditions. Genotyping using twenty-five gene-specific markers across diverse genotypes and F2 progenies revealed genes Pi5 and Pi54 to be associated with leaf blast resistance in all three populations. Genes Pib and qPbm showed an association with neck blast resistance in both natural and LR 5 × SMS populations. Additionally, a set of 184 genome-wide polymorphic markers (SSRs and SNPs), when applied to F2-resistant and F2-susceptible DNA bulks derived from LR 5 × LR 26, suggested that Pi20(t) on chromosome 12 is one of the major genes imparting disease resistance. Markers snpOS318, RM1337 and RM7102 and RM247 and snpOS316 were associated with leaf blast and neck blast resistance, respectively. The genotypes, markers and genes will help in marker-assisted selection and development of varieties with durable resistance.

QTL-Seq identified a genomic region on chromosome 1 for soil-salinity tolerance in F2 progeny of Thai salt-tolerant rice donor line "Jao Khao".

Owing to advances in high-throughput genome sequencing, QTL-Seq mapping of salt tolerance traits is a major platform for identifying soil-salinity tolerance QTLs to accelerate marker-assisted selection for salt-tolerant rice varieties. We performed QTL-BSA-Seq in the seedling stage of rice from a genetic cross of the extreme salt-sensitive variety, IR29, and "Jao Khao" (JK), a Thai salt-tolerant variety. A total of 462 F2 progeny grown in soil and treated with 160 mM NaCl were used as the QTL mapping population. Two high- and low-bulk sets, based on cell membrane stability (CMS) and tiller number at the recovery stage (TN), were equally sampled. The genomes of each pool were sequenced, and statistical significance of QTL was calculated using QTLseq and G prime (G') analysis, which is based on calculating the allele frequency differences or Δ(SNP index). Both methods detected the overlapping interval region, wherein CMS-bulk was mapped at two loci in the 38.41-38.85 Mb region with 336 SNPs on chromosome 1 (qCMS1) and the 26.13-26.80 Mb region with 1,011 SNPs on chromosome 3 (qCMS3); the Δ(SNP index) peaks were -0.2709 and 0.3127, respectively. TN-bulk was mapped at only one locus in the overlapping 38.26-38.95 Mb region on chromosome 1 with 575 SNPs (qTN1) and a Δ(SNP index) peak of -0.3544. These identified QTLs in two different genetic backgrounds of segregating populations derived from JK were validated. The results confirmed the colocalization of the qCMS1 and qTN1 traits on chromosome 1. Based on the CMS trait, qCMS1/qTN1 stably expressed 6%-18% of the phenotypic variance in the two validation populations, while qCMS1/qTN1 accounted for 16%-20% of the phenotypic variance in one validation population based on the TN trait. The findings confirm that the CMS and TN traits are tightly linked to the long arm of chromosome 1 rather than to chromosome 3. The validated qCMS-TN1 QTL can be used for gene/QTL pyramiding in marker-assisted selection to expedite breeding for salt resistance in rice at the seedling stage.

Transgenerational effects of the levonorgestrel-based birth control pill in zebrafish offspring

The consumption of hormone-derived medicines, such as levonorgestrel (LNG), is increasing worldwide, and its discharge into the environment reaches non-target organisms. In our previous study, we exposed the parental generation of zebrafish to environmentally relevant concentrations of LNG during the developmental phase. Subsequently, they had grown in a tank with clean water until adulthood. Now, we allowed this parental generation to reproduce to obtain F1 progeny unexposed to LGN, in order to analyze the transgenerational effects of parental LNG exposure on the survival and hatching of unexposed F1 embryos and the stress and behavior of F1 larvae. Here, we found decreased survival rates with higher LNG concentrations, providing a transgenerational effect. This highlights the environmental impact of exposure to LNG, causing damage at the individual and population level and affecting the next generation at the beginning of development, impacting qualities in the survival of the species.

Stable germline transgenesis using the Minos Tc1/mariner element in the sea urchin Lytechinus pictus.

Stable transgenesis is a transformative tool in model organism biology. Although the sea urchin is one of the oldest animal models in cell and developmental biology, studies in this animal have largely relied on transient manipulation of wild animals, without a strategy for stable transgenesis. Here, we build on recent progress to develop a more genetically tractable sea urchin species, Lytechinus pictus, and establish a robust transgene integration method. Three commonly used transposons (Minos, Tol2 and piggyBac) were tested for non-autonomous transposition, using plasmids containing a polyubiquitin promoter upstream of a H2B-mCerulean nuclear marker. Minos was the only transposable element that resulted in significant expression beyond metamorphosis. F0 animals were raised to sexual maturity, and spawned to determine germline integration and transgene inheritance frequency, and to characterize expression patterns of the transgene in F1 progeny. The results demonstrate transgene transmission through the germline, the first example of a germline transgenic sea urchin and, indeed, of any echinoderm. This milestone paves the way for the generation of diverse transgenic resources that will dramatically enhance the utility, reproducibility and efficiency of sea urchin research.

High-Density Genetic Map Construction and Quantitative Trait Locus Analysis of Fruit- and Oil-Related Traits in Camellia oleifera Based on Double Digest Restriction Site-Associated DNA Sequencing.

Camellia oleifera, an important tree species and source of edible oil in China, has received significant attention owing to the oil's high unsaturated fatty acid content, which has benefits for human health. However, the mechanisms underlying C. oleifera yield and oil quality are largely unknown. In this study, 180 F1 progenies were obtained from two parents with obvious differences in fruit- and oil-related traits. We constructed a high-density genetic map using a double digest restriction site-associated DNA sequencing (ddRAD-Seq) strategy in C. oleifera. This map spanned 3327 cM and anchored 2780 markers in 15 linkage groups (LGs), with an average marker interval of 1.20 cM. A total of 221 quantitative trait loci (QTLs) associated with fruit- and oil-related traits were identified across three years' worth of phenotypic data. Nine QTLs were detected simultaneously in at least two different years, located on LG02, LG04, LG05, LG06, and LG11, and explained 8.5-16.6% of the phenotypic variation in the corresponding traits, respectively. Seventeen major QTLs were obtained that explained 13.0-16.6% of the phenotypic variance. Eleven and five flanking SNPs of major QTLs for fruit- and oil-related traits were detected which could be used for marker-assisted selection in C. oleifera breeding programs. Furthermore, 202 potential candidate genes in QTL regions were identified based on the collinearity of the genetic map and the C. oleifera "CON" genome. A potential regulatory network controlling fruit development and oil biosynthesis was constructed to dissect the complex mechanism of oil accumulation. The dissection of these QTLs will facilitate the gene cloning underlying lipid synthesis and increase our understanding in order to enhance C. oleifera oil yield and quality.

Crossing a CRISPR/Cas9 transgenic tomato plant with a wild-type plant yields diverse mutations in the F1 progeny.

Generating CRISPR/Cas9-mediated mutants in tomato (Solanum lycopersicum L.) involves screening shoots regenerated from cultured cells transformed with a T-DNA harboring sequences encoding Cas9 and single guide RNAs (sgRNAs). Production of transformants can be inconsistent and obtaining transformants in large numbers may be difficult, resulting in a limited variety of mutations. Here, I report a method for generating various types of mutations from one transgenic plant harboring the CRISPR/Cas9 system. In this method, a wild-type plant was crossed with a T0 biallelic mutant expressing two sgRNAs targeting the RIPENING INHIBITOR (RIN) gene, and the resulting F1 seedlings were classified using a kanamycin resistance marker on the T-DNA. Genotyping of the RIN locus revealed that kanamycin-sensitive F1 seedlings, which carried no T-DNA, always harbored the wild-type allele and a mutant allele from the transgenic parent. Kanamycin-resistant F1 seedlings, which do carry the T-DNA, harbored a variety of novel mutant alleles, but not the wild-type allele, suggesting that it was mutated during crossing. The novel mutations included one-base insertions or short deletions at each target site, or large deletions across the two target sites. This method was also successfully applied to produce mutations in Geranylgeranyl pyrophosphate synthase 2 (GGPS2). Because this method involves crossing rather than transformation, it can be readily scaled up to produce numerous novel mutations, even in plant species or cultivars for which transformation is inefficient. Therefore, when initial transgene experiments fail to induce the desired mutation, this method provides additional opportunities for generating mutants.