Fertility Regulation Research Articles

Cellular atlases of ovarian microenvironment alterations by diet and genetically-induced obesity.

Obesity, which can arise from genetic or environmental factors, has been shown to cause serious damages to the reproductive system. The ovary, as one of the primary regulators of female fertility, is a complex organ comprised of heterogeneous cell types that work together to maintain a normal ovarian microenvironment (OME). Despite its importance, the effect of obesity on the entire ovary remains poorly documented. In this study, we performed ovary single-cell and nanoscale spatial RNA sequencing to investigate how the OME changed under different kinds of obesity, including high-fat diet (HFD) induced obesity and Leptin ablation induced obesity (OB). Our results demonstrate that OB, but not HFD, dramatically altered the proportion of ovarian granulosa cells, theca-interstitial cells, luteal cells, and endothelial cells. Furthermore, based on the spatial dynamics of follicular development, we defined four subpopulations of granulosa cell and found that obesity drastically disrupted the differentiation of mural granulosa cells from small to large antral follicles. Functionally, HFD enhanced follicle-stimulating hormone (FSH) sensitivity and hormone conversion, while OB caused decreased sensitivity, inadequate steroid hormone conversion, and impaired follicular development. These differences can be explained by the differential expression pattern of the transcription factor Foxo1. Overall, our study provides a powerful and high-resolution resource for profiling obesity-induced OME and offers insights into the diverse effects of obesity on female reproductive disorders.

The Neurod1/4-Ntrk3-Src pathway regulates gonadotrope cell adhesion and motility

Pituitary gonadotrope cells are essential for the endocrine regulation of reproduction in vertebrates. These cells emerge early during embryogenesis, colonize the pituitary glands and organize in tridimensional networks, which are believed to be crucial to ensure proper regulation of fertility. However, the molecular mechanisms regulating the organization of gonadotrope cell population during embryogenesis remain poorly understood. In this work, we characterized the target genes of NEUROD1 and NEUROD4 transcription factors in the immature gonadotrope αT3-1 cell model by in silico functional genomic analyses. We demonstrated that NEUROD1/4 regulate genes belonging to the focal adhesion pathway. Using CRISPR/Cas9 knock-out approaches, we established a double NEUROD1/4 knock-out αT3-1 cell model and demonstrated that NEUROD1/4 regulate cell adhesion and cell motility. We then characterized, by immuno-fluorescence, focal adhesion number and signaling in the context of NEUROD1/4 insufficiency. We demonstrated that NEUROD1/4 knock-out leads to an increase in the number of focal adhesions associated with signaling abnormalities implicating the c-Src kinase. We further showed that the neurotrophin tyrosine kinase receptor 3 NTRK3, a target of NEUROD1/4, interacts physically with c-Src. Furthermore, using motility rescue experiments and time-lapse video microscopy, we demonstrated that NTRK3 is a major regulator of gonadotrope cell motility. Finally, using a Ntrk3 knock-out mouse model, we showed that NTRK3 regulates gonadotrope cells positioning in the developing pituitary, in vivo. Altogether our study demonstrates that the Neurod1/4-Ntrk3-cSrc pathway is a major actor of gonadotrope cell mobility, and thus provides new insights in the regulation of gonadotrope cell organization within the pituitary gland.

In Vitro Study on Spermicidal Action of Hydro-methanol Extract of Tinospora cordifolia (Willd.) Stem in Rat and Human Sperm: a Comparative Analysis.

Human fertility regulation is a major way to control overpopulation. In this perspective, this study emphasized the in vitro effect of hydro-methanol extract of Tinospora cordifolia (TCHME) stem for spermicidal and reproductive hypo-functions using human and rat samples. Control, 0.5-, 1-, and 2-mg TCHME-charged groups were considered to assess the relevant parameters. Levels of spermiological parameters like sperm motility, viability, the integrity of plasma and acrosomal membrane, and nuclear chromatin decondensation were significantly reduced (p < 0.05) in the dose- and duration-dependent TCHME-charged groups compared to the control. The inhibitory concentration 50 (IC50) of TCHME on motile human and rat sperms were 0.8 and 0.4 mg/ml, respectively. Testicular androgenic key enzymes and antioxidant enzymes (human sperm pellet, testes, and epididymis of rat)' activities were significantly diminished (p < 0.05), while antioxidant enzymes' activities were significantly elevated (p < 0.05) in renal and insignificantly (p > 0.05) elevated in hepatic tissues of rat in TCHME-charged groups compared to the control. Significant elevation (p < 0.05) of thiobarbituric acid reactive substances (TBARS)' level in human sperm pellet, testes, and epididymis of rats and significant diminution (p < 0.05) in TBARS levels of liver and kidney were observed in TCHME-charged groups. It focused that TCHME is more potent for stress imposition on reproductive tissues and sperm compared to the other tested tissues. Non-significant alterations (p > 0.05) in glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT) activities in the said organs of rat indicated its non-toxic effect. It highlighted that TCHME possesses spermicidal and reproductive tissue-specific effects which strengthen the possibilities of male contraceptive development from it.

Litterfall and nutrient transfer dynamics in a successional gradient of tropical dry forest in Colombia

Introduction: The litterfall production, foliar nutrient dynamics and decomposition are essential to maintain nutrient cycling, soil fertility, and carbon regulation in terrestrial ecosystems. With several studies addressing the variation of these processes, their dynamics in tropical dry forests (TDFs) remain unclear, due to its complex interaction of biotic and abiotic factors. Objective: To evaluate litterfall, nutrient potential return and use efficiency, and decomposition variation in a TDF successional gradient in Tolima, Colombia. Methods: We quantified litterfall from November 2017 to October 2019 in 12 plots distributed in four successional stages: initial, early, intermediate, and late forests. We identified key tree species in foliar litter production and characterized the foliar decomposition of these species. At the community level, we quantified the C, N and P potential return, the N and P use efficiency, and the C:N and N:P ratio. Subsequently, we analyze relationships between vegetation characteristics and some soil chemical properties with these ecological processes. Results: We found that total litterfall in late forests (8.46 Mg ha-1 y-1) was double that found in initial forests (4.45 Mg ha-1 y-1). Decomposition was higher in initial (k = 1.28) compared to intermediate (k = 0.97) and late forests (k = 0.87). The nutrient potential return didn’t change along succession, but it did show differences between study sites. The structural development and species richness favored litterfall, while soil chemical conditions influenced nutrient returns and decomposition. Conclusions: TDFs could recover key ecosystem function related to litterfall and nutrient dynamics after disturbances cessation; however, the soil quality is fundamental in return and release of nutrients.

Validation of a new Custom Polyclonal Progesterone Receptor Antibody for Immunohistochemistry in the Female Mouse Brain.

Immunohistochemical visualization of progesterone receptor (PR)-expressing cells in the brain is a powerful technique to investigate the role of progesterone in the neuroendocrine regulation of fertility. A major obstacle to the immunohistochemical visualization of progesterone-sensitive cells in the rodent brain has been the discontinuation of the commercially produced A0098 rabbit polyclonal PR antibody by DAKO. To address the unavailability of this widely used PR antibody, we optimized and evaluated 4 alternative commercial PR antibodies and found that each lacked the specificity and/or sensitivity to immunohistochemically label PR-expressing cells in paraformaldehyde-fixed female mouse brain sections. As a result, we developed and validated a new custom RC269 PR antibody, directed against the same 533-547 amino acid sequence of the human PR as the discontinued A0098 DAKO PR antibody. Immunohistochemical application of the RC269 PR antibody on paraformaldehyde-fixed mouse brain sections resulted in nuclear PR labeling that was highly distinguishable from background, specific to its antigen, highly regulated by estradiol, matched the known distribution of PR protein expression in the female mouse hypothalamus, and nearly identical to that of the discontinued A0098 DAKO PR antibody. In summary, the RC269 PR antibody is a specific and sensitive antibody to immunohistochemically visualize PR-expressing cells in the mouse brain.

Mapping soil organic matter and identifying potential controls in the farmland of Southern China: Integration of multi‐source data, machine learning and geostatistics

AbstractSoil organic matter (SOM) plays a critical role in terrestrial ecosystem functioning and is closely related to many global issues like soil fertility, soil health and climate regulation. Therefore, obtaining accurate information on the spatial distribution of SOM and its potential controlling factors is of global interest. However, this remains a great challenge since SOM is affected by numerous natural and anthropogenic factors and usually showed strong heterogeneity. In this study, we collected a total of 16,580 surface soil (0–20 cm) samples from the farmland throughout Jiangxi Province. And the Random Forest (RF), Cubist and gradient‐boosted models were compared and used to define the factor which is most associated with SOM. Then the ordinary kriging (OK) and machine learning‐ordinary co‐kriging (ML‐COK) were used to map SOM. We found that on average, 30.86 g kg−1 SOM was present in farmland soil of Jiangxi Province. Anthropogenic activities strongly affected SOM level, with five of the top 10 most important factors are anthropogenic related. The straw return amount was proved to have the largest importance (31.46%) for modelling SOM and a significant (p &lt; 0.001) positive relationship between SOM content and the amount of straw returned to farmland was detected. Additionally, returning straw improved crop production. Soil derived from the Quaternary Subred Sand has the highest SOM content (37.82 g kg−1). Crop rotation also improved SOM content and the rice‐bean rotation system has the highest SOM content (34.27 g kg−1). With the best performance, the RF algorithm (R2 = 0.49, RMSE = 6.77 g kg−1) was selected to identify the primary control of SOM and integrated with COK, which we termed as ML‐COK, to map the SOM in the farmland of Jiangxi Province. ML‐COK outperformed OK method for mapping the SOM in farmland of Jiangxi Province with R2 of 0.351 and Lin's concordance correlation coefficient of 0.549. Farmland distributed in the central part of the province had high SOM content. In contrast, farmland in the north, south and east parts had relatively low SOM. Our study offers new insight for mapping soil properties, identifying potential factors driving variation in SOM, and also provides valuable information for making more reasonable and environmentally friendly farmland management measures.

Histone H3 lysine 27 trimethylation suppresses jasmonate biosynthesis and signaling to affect male fertility under high temperature in cotton

High-temperature (HT) stress causes male sterility in crops, thus decreasing yields. To explore the possible contribution of histone modifications to male fertility under HT conditions, we defined the histone methylation landscape for the marks histone H3 lysine 27 trimethylation (H3K27me3) and histone H3 lysine 4 trimethylation (H3K4me3) by chromatin immunoprecipitation sequencing (ChIP-seq) in two differing upland cotton (Gossypium hirsutum) varieties. We observed a global disruption in H3K4me3 and H3K27me3 modifications, especially H3K27me3, in cotton anthers subjected to HT. HT affected the bivalent H3K4me3–H3K27me3 modification more than either monovalent modification. We determined that removal of H3K27me3 at the promoters of jasmonate-related genes increased their expression, maintaining male fertility under HT in the HT-tolerant variety at the anther dehiscence stage. Modulating jasmonate homeostasis or signaling resulted in an anther indehiscence phenotype under HT. Chemical suppression of H3K27me3 deposition increased jasmonic acid contents and maintained male fertility under HT. In summary, our study provides new insights into the regulation of male fertility by histone modifications under HT and suggests a potential strategy for improving cotton HT tolerance.

Effects of follicle-stimulating hormone, insulin-like growth factor 1, fibroblast growth factor 2, and fibroblast growth factor 9 on sirtuins expression and histone deacetylase activity in bovine granulosa cells

Granulosa cells (GC) are critical regulators of fertility. During the process of ovarian folliculogenesis, these cells undergo profound changes while producing steroid hormones that are important to control follicular growth, oocyte maturation, and ovulation. Sirtuins are enzymes that regulate several biological processes and have been associated with control of GC function. However, how sirtuins are regulated in GC during ovarian folliculogenesis remains to be unveiled. The present study was designed to investigate effects of hormones that control GC proliferation, differentiation, and steroidogenesis on expression of the seven members of the mammalian sirtuins family (SIRT1-7) and on histone deacetylase activity of nuclear sirtuins (SIRT1, 6, and 7) in GC. Bovine granulosa cells were isolated from small antral follicles (1–5 mm) and were treated with or without follicle-stimulating hormone (FSH), insulin-like growth factor 1 (IGF-1), and fibroblast growth factors 2 (FGF2) and 9 (FGF9). Following treatments, cell proliferation was determined via a cell analyzer, estradiol synthesis and histone deacetylase activity were determined via ELISA, and sirtuins mRNA expression was determined via qPCR. Treatments with FSH and IGF-1 stimulated cell proliferation while addition of FGF2 or FGF9 suppressed estradiol production stimulated by FSH plus IGF-1. In terms of treatments that regulated sirtuins expression in GC, fibroblast growth factors were the most impactful: FGF2 alone increased SIRT1 mRNA expression in comparison to several treatments and increased mRNA abundance of SIRT2 and SIRT7 when added to the combination of FSH and IGF-1; the addition of FGF9 to the combination of FSH and IGF-1 increased mRNA expression of SIRT2, SIRT3, SIRT4, SIRT6, and SIRT7 and increased mRNA expression of SIRT5 in comparison to the negative control group that received no treatment. Also, FGF2 alone increased histone deacetylase activity of sirtuins in comparison to all treatments that contained FSH and/or IGF-1. Furthermore, several correlations were observed between treatments and sirtuins expression and activity, between estradiol or GC numbers and sirtuins expression, and between expression of sirtuins. As FGF2 and FGF9 are considered anti-differentiation factors of GC that stimulate GC proliferation while suppressing estradiol production in combination with FSH and IGF-1, data of this study suggest that sirtuins are associated with control of differentiation of bovine GC.

O-244 Mitochondrial fission factor (MFF) is required for female fertility and oocyte development

Abstract Study question To explore the role of Mitochondrial fission factor (MFF) in female fertility and oocyte development using a Mff knockout mouse model. Summary answer Mff deficiency influences mitochondrial dynamics in oocytes at germinal vesicle stage (GV), and results in premature ovarian failure (POF) and female infertility. What is known already Mff regulates mitochondrial quality control (MQC) by influencing mitochondrial dynamics. MQC has critical effects on oocyte development and POF. However, little is known about the role of Mff in oocyte development and female fertility. Study design, size, duration The Mff knockout (Mff-/-) mice were generated via the CRISPR-Cas9 system. Adult Mff-/- female mice were compared to wildtype (WT) female mice. Female mice at 2-month old from each group (Mff-/- or WT, n = 5) were used to assess the fecundity for 10 months. The follicle development (n = 4 mice), the number of GV oocytes (n = 5 mice) and metaphase II (MII) oocytes (n = 5 mice) were determined. Participants/materials, setting, methods Female mice from each group were mated with adult WT males to assess the fecundity. Follicle development was evaluated in ovaries after fixation, paraffin embedding, and sectioning, followed by hematoxylin and eosin staining. MFF and DRP1 protein expression was evaluated by western blot. Oocyte mitochondria were stained with the Mito-Tracker Red CMXRos. Main results and the role of chance The Mff deficient ovaries had significantly lower levels of MFF (0.056 vs. 1, p &amp;lt; 0.01) than the WT ovaries. Mff-/- female mice were infertile (0 vs. 39, p &amp;lt; 0.05). Mature (8-week-old) Mff deficient ovaries had significantly lower weight (0.0043 vs. 0.011, p &amp;lt; 0.05) compared to the WT. The 2-month-old female Mff-/- mice had a lower number of primordial (220 vs. 775, p&amp;lt; 0.05), primary (128 vs. 580, p&amp;lt; 0.05), secondary (29 vs. 82, p&amp;lt; 0.05) and antral follicles (5 vs. 23, p&amp;lt; 0.05). The numbers of GV oocytes (18 vs. 37, p&amp;lt; 0.05) and MII oocytes (2 vs. 16, p&amp;lt; 0.001) were lower in the Mff-/- mice than the WT mice. In addition, the Mff-/- ovaries expressed significantly less mitochondrial fission key regulator DRP1 (0.54 vs. 1, p&amp;lt; 0.05). Mff-/- oocytes contained elongated and aggregated mitochondria compared to the discrete and evenly distributed mitochondria in the ooplasm of WT oocytes. Herein, we found that the Mff played a concerted role in oocyte and follicle development. Limitations, reasons for caution The present observations in mice may not be applicable to humans. The results are derived global knockout of Mff. It is not known which kind of cell plays a vital role in fertility. Further investigation is needed to explore the regulatory mechanism of Mff in oocyte development and fertility regulation. Wider implications of the findings This study provides important insights into the relationship between mitochondrial dynamics and female fertility. Trial registration number not applicable

P-650 Follicular extracellular vesicles of older women inhibit oocyte maturation

Abstract Study question Whether follicular extracellular vesicles of older women interfere with the quality of oocytes. Summary answer OLD-EVs induced the oxidative stress in the oocytes and inhibited oocyte maturation by increasing the abnormal mitochondria distribution and abnormal spindle rates. What is known already Ovary is a critical regulator of female fertility and the source of oocytes. With the modern tendency of delaying childbearing, ovarian aging has become an age-related disease. Ovarian aging is generally characterized by a gradual decrease in both the quantity and quality of oocytes. The pregnancy outcomes of older women were far from satisfying with higher abortion rates, birth defects rates, and lower live birth rates even after the assistance of IVF. However, the detailed mechanisms of decreased oocyte quality of aging women have not been clarified. Study design, size, duration Extracellular vesicles (EVs) were isolated from follicular fluid of older women (aged 40-50, n = 7, OLD-EVs) and younger women (aged 25-30, n = 6, YOUNG-EVs) via ultracentrifugation. Germinal vesicle (GV) oocytes from female ICR mice were co-cultured with OLD-EVs, YOUNG-EVs, or PBS (blank control), respectively. GV breakdown (GVBD) rate and maturation rate were calculated at two-hour and fourteen-hour of co-culture, respectively. Besides, oocyte mitochondria distribution, meiosis spindle morphology, and oxidative status were assessed in different groups. Participants/materials, setting, methods EVs were determined by western blotting, nanoparticle tracking analysis, and transmission electron microscopy. Fluorescence labeled EVs were used to visualize internalization by oocytes. Oocytes mitochondria and mitosis spindles were stained with fluorescence, and abnormal mitochondria rate or abnormal spindle rate was calculated. Reactive oxygen species (ROS) level was detected in the differently treated oocytes. Moreover, the expression of CAT, GSS, and SOD was determined in the oocytes using quantitative reverse transcription polymerase chain reaction. Main results and the role of chance Both OLD-EVs and YOUNG-EVs are bilayered vesicles, ranging from 100 to 150 nm and enriched in Alix, TSG101, and CD9. EVs could be internalized by oocytes within one hour. After coculture, GVBD rate was similar among the three groups; whereas maturation rate was significantly decreased in the OLD-EV group compared with YOUNG-EV group or PBS group. In addition, the abnormal mitochondria distribution rate or abnormal spindle rate were significantly increased in the OLD-EV group compared with PBS or YOUNG-EV group. The ROS level was increased in the PCOS-EV group compared with YOUNG-EV group, and the expression of CAT, GSS, and SOD was increased in the OLD-EV-treated oocytes. Limitations, reasons for caution Our study did not identify the contents of OLD-EVs and YOUNG-EVs, and the molecular mechanisms of dysregulations induced by OLD-EVs need further researches to investigate. Wider implications of the findings This work confirmed that EV-conducted cellular communication played an important role in oocyte maturation. The dysregulation of oocytes induced by OLD-EVs might be related to the poor oocyte quality of aging women, which provide a novel target to improve pregnancy outcomes of these patients. Trial registration number not applicable

O-095 Different phenotypes of PCOS: Implications to clinical practice

Abstract Polycystic ovary syndrome (PCOS) is a very common, heterogeneous syndrome which presents with a spectrum of clinical and biochemical features. It is associated with both reproductive and metabolic dysfunction and has a negative impact on long term health, including mental health. Although there appears to be a common genetic background to PCOS, regardless of presentation, environmental factors, including diet, play an important part in determining the phenotype. The most commonly used diagnostic criteria, the Rotterdam criteria, takes account of the heterogenous nature of the syndrome but the spectrum of presentation can be categorised as four clinical and biochemical subtypes which comprise (A) ovulatory dysfunction, hyperandrogenism, polycystic ovaries (PCO), (B) ovulatory dysfunction, hyperandrogenism but without PCO, (C) hyperandrogenism without ovulatory dysfunction but with PCO and (D) ovulatory dysfunction with PCO but without hyperandrogenism. These are not rigid categories since some women may go from one to another, for example weight gain may lead to anovulation in a woman who previously had hirsutism but regular cycles. Nevertheless, the diagnostic sub-category has a significance influence on clinical management. Women who have both hyperandrogenism and ovulatory dysfunction (A&amp;B) are at higher risk of metabolic dysfunction which is exacerbated by obesity (and carries a high long-term risk of diabetes and cardiovascular disease) and so merit thorough investigation which includes assessment of glucose tolerance and lipids. Those who have predominantly ovulatory problems may require fertility treatment or regulation of menses but some who simply have irregular menses may need no specific treatment. Symptoms of hyperandrogenism (hirsutism, acne, alopecia) can be particularly distressing and may be best managed in collaboration with a dermatologist. The negative impact of PCOS on mental health is a very common problem, the high prevalence of which has only recently been appreciated. Not surprisingly, it is particularly relevant in those women who have moderate to severe symptoms of hyperandrogenism. In summary, PCOS is a syndrome characterised by a variety of symptoms and endocrine abnormalities and it is important to tailor treatment individually, according to the need for symptom relief and with regard to concerns about long-term health.

P-691 Does serum Kisspeptin levels discriminate between fertile and infertile women with polycystic ovary syndrome?

Abstract Study question Does serum Kisspeptin (KP) levels discriminate between fertile and infertile women with polycystic ovary syndrome (PCOS)? Summary answer Serum KP levels can be used as a potential discriminatory biomarker for infertility in women with PCOS. What is known already PCOS is the most common cause of anovulatory infertility and not fully elucidated pathology. A persistent rapid gonadotropin-releasing hormone (GnRH) pulse frequency in patients with PCOS exist throughout ovulatory cycle. KP is a neuropeptide that increases GnRH pulsatile release during ovulation. Therefore, KP may have a key role as a central regulator of fertility in PCOS patients. Although, recent studies showed that the KP concentration is higher in PCOS patients, the evidence is limited as to whether serum KP levels determine infertility in PCOS. Study design, size, duration This was a single center prospective cohort study conducted at Goztepe Prof. Dr. Suleyman Yalcin City Hospital Affiliated to Istanbul Medeniyet University, Istanbul, Turkey between January 2023 to February 2023. Our study enrolled 30 fertile and 30 infertile women with PCOS, who were aged between 18 and 45 years. PCOS was defined according to the criteria of the Rotterdam ESHRE- ASRM sponsored consensus group (2004). Participants/materials, setting, methods Venous blood samples from the participants were obtained after fasting and between 8 a.m. and 10 a.m. on days 2-5 of the menstrual cycle. Samples were thawed, and a human KISS1 (Kisspeptin 1) ELISA kit (Elabscience, USA,lot no:E-EL-H5618) was applied to measure KP serum concentrations. Patient characteristics and KP concentrations were compared among the two groups. Statistical analysis was performed by Mann-Whitney, Spearman correlations, and linear regression analysis. A p-value &amp;lt;0.05 was considered significant. Main results and the role of chance The mean age of the patients was 27.57±5.39 years in the fertile PCOS group and 26.80± 4.85 years in the infertile PCOS group (p = 0.63). There was no significant difference among the groups in terms of body mass index (BMI), duration of infertility, serum FSH, LH, LH-to-FSH ratio, E2, antimullerian hormone (AMH), TSH, prolactin, 17OHP, total testosterone, SHBG, HbA1c, HOMA-IR, neutrophil–lymphocyte ratio levels and antral follicle count (AFC). DHEA-S was significantly higher in the infertile PCOS group (306.73±94.869 mcg/ dL), compared to the fertile PCOS group (258.08±84.29 mcg/ dL, p = 0.037). The mean KP level was significantly higher in the infertile PCOS group (444.80±136.61 ng/ mL), compared to the fertile PCOS group (333.02±131.10 ng/ mL, p = 0.001). The KP level was positively correlated with AFC, AMH, and total testosterone levels (R = 0.495, R = 0.548, R = 0.362, p &amp;lt; 0.05, respectively) in the infertile PCOS group. Furthermore, ROC analysis showed that the optimal cut-point value was found to be 285.59. When this cut-off value of serum KP levels was taken, the sensitivity was 0.96 and the specificity was 0.50. Linear regression analysis revealed that AMH was positively associated with KP levels (p = 0.022). Limitations, reasons for caution Due to the limited number of women with available samples, we were not able to analyze KP serum levels according to specific PCOS phenotypes Wider implications of the findings Serum KP levels are higher in infertile women with PCOS compared to fertile women with PCOS. Infertility caused by PCOS can be predicted by serum KP levels. In the future identification, KP in PCOS may be applied to develop potential therapeutic agents. Trial registration number Not applicable

The NAC transcription factor ZmNAC132 regulates leaf senescence and male fertility in maize

Leaf senescence is an integral step in the final stages of plant development, as nutrient remobilization from leaves to sink organs is accomplished during this process. NACs compose a large superfamily of plant-specific TFs involved in multiple plant development processes. Here, we identified a maize NAC TF, ZmNAC132, involved in leaf senescence and male fertility. ZmNAC132 expression was tightly linked to leaf senescence in an age-dependent manner. Knockout of ZmNAC132 led to delays in chlorophyll degradation and leaf senescence, whereas overexpression of ZmNAC132 had the opposite effects. ZmNAC132 could bind to and transactivate the promoter of ZmNYE1, a major chlorophyll catabolic gene, to accelerate chlorophyll degradation during leaf senescence. Moreover, ZmNAC132 affected male fertility through the upregulation of ZmEXPB1, an expansin-encoding gene associated with sexual reproduction and other related genes. Together, the results show that ZmNAC132 participates in the regulation of leaf senescence and male fertility through the targeting of different downstream genes in maize.

Gibberellin confers to the expression of TaGA-6D and negatively regulates the fertility of wheat with Aegilops juvenalis cytoplasm

As the trace signal molecules widely existing in plants, plant hormones can regulate physiological responses of plants at low concentrations. At present, the effect of plant endogenous hormones on wheat male fertility has attracted attention, but the molecular mechanism underlying fertility regulation is unclear. Given this, the anthers of five isonuclear alloplasmic male sterile lines and their maintainer line were RNA-sequenced. A gene TaGA-6D encoding gibberellin (GA) regulated protein was isolated, which was located to the nucleus, cell wall and/or cell membrane, and predominantly highly expressed in the anther of Ju706A, a male sterile line with Aegilops juvenalis cytoplasm. By spraying assay of GA with different concentrations on fertility line Ju706R, it was found that with the increase of exogenous GA concentration, the content of endogenous GA and expression level of TaGA-6D in anther gradually increased, and the fertility decreased. However, silencing of TaGA-6D partially restore the fertility of Ju706R sprayed with 1000 ng/μl GA, and indicating that gibberellin can promote the expression of TaGA-6D and negatively regulates the fertility of wheat with Aegilops juvenalis cytoplasm, which providing new insights for understanding hormone regulation of male fertility in wheat.

Amino acid fertilizer strengthens its effect on crop yield and quality by recruiting beneficial rhizosphere microbes.

There is currently an increase in the use of new types of fertilizers in modern agriculture. Studies have shown that amino acid fertilizers can improve crop yield and quality. However, their effects on crop rhizosphere ecology and their ecological impacts on crop yield are largely unknown. This study evaluated the effects of a water-soluble amino acid fertilizer (WAAF) on tomatoes and its ecological effects on rhizosphere bacterial communities using greenhouse pot experiments. The results showed that WAAF could promote the growth of tomatoes and improve the quality of fruits more effectively than water-soluble chemical fertilizer controls. Interestingly, WAAF showed a different regulating pattern on root exudates and increased the secretion of 17 major water-soluble root exudates, including hexadecanoic acid and 3-hydroxy-γ-butyrolactone. Water-soluble amino acid fertilizer also affected noticeably the composition, abundance, and beta-diversity of rhizosphere bacterial communities, and strengthened the potential relationships between community members. Water-soluble amino acid fertilizer showed a significant selective enrichment ability and recruited some members of the genera such as Cupriavidus, Ralstonia, Chitinophaga, Gemmatimonas, Mitsuaria, Mucilaginibacter, Paracoccus, Sphingopyxis, and Variovorax. Network analysis and functional prediction implied that, besides fertilizer effects, the recruiting of beneficial microbes involved in chemotaxis and biofilm formation was also a considerable factor in tomato yield and quality improvement. Our study revealed ecological and recruiting effects of WAAF on rhizosphere microbes and potentially beneficial microbiota, and provided a basis for the amino acid fertilizer regulation of rhizosphere ecology to improve soil health and further improve crop yield and quality. © 2023 Society of Chemical Industry.

Network interpenetrating slow-release nitrogen fertilizer based on carrageenan and urea: A new low-cost water and fertilizer regulation carrier

Modern agriculture presents new requirements of low cost, high water retention and degradability for superabsorbent and slow-release fertilizers. In this study, carrageenan (CG), acrylic acid (AA), N, N ‘-methylene diacrylamide (MBA), urea and ammonium persulfate (APS) were used as raw materials. A kind of high water absorption, water retention, nitrogen slow release and biodegradable carrageenan superabsorbent (CG-SA) was prepared by grafting copolymerization. The optimal CG-SA was obtained with a water absorption rate of 680.45 g/g by orthogonal L18(3)7 experiments and single-factor experiments. The water absorption behavior of CG-SA in deionized water and salt solution were studied. The CG-SA was characterized before and after degradation by FTIR, SEM. The nitrogen release behavior and kinetic characteristics of CG-SA were investigated. In addition, CG-SA degraded 58.33 % and 64.35 % in soil at 25 °C and 35 °C after 28 days. All the results indicated that the low-cost and degradable CG-SA can achieve simultaneous slow release of water and nutrients, which is expected to be widely used as a new water-fertilizer integration technology in arid and poor areas.

Identifying new receptors in the regulation of fertility

Searchable abstracts of presentations at key conferences in endocrinology ISSN 1470-3947 (print) | ISSN 1479-6848 (online)

DOT1L promotes spermatid differentiation by regulating expression of genes required for histone-to-protamine replacement.

Unique chromatin remodeling factors orchestrate dramatic changes in nuclear morphology during differentiation of the mature sperm head. A crucial step in this process is histone-to-protamine exchange, which must be executed correctly to avoid sperm DNA damage, embryonic lethality and male sterility. Here, we define an essential role for the histone methyltransferase DOT1L in the histone-to-protamine transition. We show that DOT1L is abundantly expressed in mouse meiotic and postmeiotic germ cells, and that methylation of histone H3 lysine 79 (H3K79), the modification catalyzed by DOT1L, is enriched in developing spermatids in the initial stages of histone replacement. Elongating spermatids lacking DOT1L fail to fully replace histones and exhibit aberrant protamine recruitment, resulting in deformed sperm heads and male sterility. Loss of DOT1L results in transcriptional dysregulation coinciding with the onset of histone replacement and affecting genes required for histone-to-protamine exchange. DOT1L also deposits H3K79me2 and promotes accumulation of elongating RNA Polymerase II at the testis-specific bromodomain gene Brdt. Together, our results indicate that DOT1L is an important mediator of transcription during spermatid differentiation and an indispensable regulator of male fertility.

Constraints using the liquid fraction from roadside grass as a bio‐based fertilizer

AbstractBackgroundRoadside grass cuttings are currently considered a waste product due to their association with road sweepings as contaminated waste, therefore, their potential as a biofertilizer is understudied.AimThis study aimed to determine whether grass liquid fraction (GLF) collected from a roadside verge in Maldegem, Belgium, and pressed using a screw press was suitable as a biofertilizer.MethodsThe characterization of the heavy metal content of the GLF was conducted using an ICP‐OES. From May to September 2019, a pot experiment was set up using a randomized block design to compare tomato plant growth, yield, and nutrition for GLF‐treated plants to two commercial fertilizers and tap water as a control.ResultsThe heavy metal content of the GLF was below the maximum permissible concentrations (MPCs) for organic fertilizers as set out by the European Comission fertilizer regulation 1069/2009 and 1107/2009 (European Comission, 2019). However, despite having a fairly well‐balanced nutrient content (0.1% N, 0.04% P2O5, and 0.2% K2O), GLF had a negative effect on the growth, root weight, and yield of the tomato plants, killing six out of ten plants. GLF also promoted mold growth in the soil of some plants. Since the GLF was uncontaminated, heavy metal toxicity did not cause the negative effect.ConclusionsPrevious research showed that liquid fractions from some plants negatively affect the growth of others due to allelopathic chemicals; this, together with the stimulation of fungal growth, could have caused the negative effects observed. Future experiments will investigate the herbicidal property of GLF and possible treatments to potentially recover the nutrients contained within the GLF for application as a biofertilizer.

The miR156b-GmSPL2b module mediates male fertility regulation of cytoplasmic male sterility-based restorer line under high-temperature stress in soybean.

High-temperature (HT) stress at flowering stage causes significant damage to soybean, including pollen abortion and fertilization failure, but few genes involved in male fertility regulation under HT stress in soybean have been characterized. Here, we demonstrated that miR156b-GmSPL2b module involved in male fertility regulation of soybean cytoplasmic male sterility (CMS)-based restorer line under HT stress. Overexpression of miR156b decreased male fertility in soybean CMS-based restorer line and its hybrid F1 with CMS line under HT stress. RNA-seq analysis found that miR156b mediated male fertility regulation in soybean under HT stress by regulating the expression of pollen development and HT response related genes. Metabolomic analysis of miR156bOE revealed reduction in flavonoid content under HT stress. Integrated transcriptomic and metabolomic analysis showed that the overexpression of miR156b caused flavonoid metabolism disorder in soybean flower bud under HT stress. Knockout of GmSPL2b also decreased the thermotolerance of soybean CMS-based restorer line during flowering. Moreover, GmSPL2b turned out to be directly bounded to the promoter of GmHSFA6b. Further verification indicated that GmHSFA6b overexpression enhanced HT tolerance in Arabidopsis during flowering. Substance content and gene expression analysis revealed that miR156b-GmSPL2b may mediate reactive oxygen species clearance by regulating flavonoid metabolism, thus participating in the regulation of male fertility in soybean under HT stress. This study not only provided important progress for understanding the molecular mechanism of miR156b-GmSPL2b regulating the male fertility of soybean CMS-based restorer line under HT stress, but also provided genetic resources and theoretical basis for creating HT-tolerant strong restorer lines.