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TTC12 is a cytoplasmic and centromere-localized protein that plays a role in the proper assembly of dynein arm complexes in motile cilia in both respiratory cells and sperm flagella. This finding underscores its significance in cellular motility and function. However, the wide role of TTC12 in human spermatogenesis-associated primary ciliary dyskinesia (PCD) still needs to be elucidated. Whole-exome sequencing (WES) and Sanger sequencing were performed to identify potentially pathogenic variants causing PCD and multiple morphological abnormalities of sperm flagella (MMAF) in an infertile Pakistani man. Diagnostic imaging techniques were used for PCD screening in the patient. Real-time polymerase chain reaction (RT‒PCR) was performed to detect the effect of mutations on the mRNA abundance of the affected genes. Papanicolaou staining and scanning electron microscopy (SEM) were carried out to examine sperm morphology. Transmission electron microscopy (TEM) was performed to examine the ultrastructure of the sperm flagella, and the results were confirmed by immunofluorescence staining. Using WES and Sanger sequencing, a novel homozygous missense variant (c.C1069T; p.Arg357Trp) in TTC12 was identified in a patient from a consanguineous family. A computed tomography scan of the paranasal sinuses confirmed the symptoms of the PCD. RT–PCR showed a decrease in TTC12 mRNA in the patient's sperm sample. Papanicolaou staining, SEM, and TEM analysis revealed a significant change in shape and a disorganized axonemal structure in the sperm flagella of the patient. Immunostaining assays revealed that TTC12 is distributed throughout the flagella and is predominantly concentrated in the midpiece in normal spermatozoa. In contrast, spermatozoa from patient deficient in TTC12 showed minimal staining intensity for TTC12 or DNAH17 (outer dynein arms components). This could lead to MMAF and result in male infertility. This novel TTC12 variant not only illuminates the underlying genetic causes of male infertility but also paves the way for potential treatments targeting these genetic factors. This study represents a significant advancement in understanding the genetic basis of PCD-related infertility. [ABSTRACT FROM AUTHOR]

Four composting experiments were adopted to evaluate the effect of inoculating mature compost on the harmless treatment of avermectin fermentation residue (AFR): added 100% spent mushroom substrate (SMS) and no AFR in CK, 15% AFR and 85% SMS in T1, 30% AFR and 70% SMS in T2, 30% AFR, 60% SMS and 10% mature compost in T3. Results showed that, compared with that of T2, the thermophilic stage in T3 arrived 12 days earlier and extended by 5 days, the GI reached above 90% 6 days earlier, and the changes of pH, EC, nitrate nitrogen, ammonium nitrogen and C/N ratio were all correspondingly advanced. The number of total OTUs and unique OTUs were increased, the dominant microbial populations and the biodegradation of avermectin were improved. Inoculating mature compost also had a significant impact on the expression of antibiotic resistance genes, as it significantly improved the bacterial community of compost. The addition of AFR was increased from 15 to 30% without delaying the composting process, which was conducive to the harmless treatment of AFR, and was of great significance to the cleaner production of avermectin factories. [ABSTRACT FROM AUTHOR]

Polyploidization is a process which is related to species hybridization and whole genome duplication. It is widespread among angiosperm evolution and is essential for speciation and diversification. Allopolyploidization is mainly derived from interspecific hybridization and is believed to pose chromosome imbalances and genome instability caused by meiotic irregularity. However, the self-compatible allopolyploid in wild nature is cytogenetically and genetically stable. Whether this stabilization form was achieved in initial generation or a consequence of long term of evolution was largely unknown. Here, we synthesized a series of nascent allotetraploid wheat derived from three diploid genomes of A, S*, and D. The chromosome numbers of the majority of the progeny derived from these newly formed allotetraploid wheat plants were found to be relatively consistent, with each genome containing 14 chromosomes. In meiosis, bivalent was the majority of the chromosome configuration in metaphase I which supports the stable chromosome number inheritance in the nascent allotetraploid. These findings suggest that diploidization occurred in the newly formed synthetic allotetraploid wheat. However, we still detected aneuploids in a proportion of newly formed allotetraploid wheat, and meiosis of these materials present more irregular chromosome behavior than the euploid. We found that centromere pairing and centromere clustering in meiosis was affected in the aneuploids, which suggest that aneuploidy may trigger the irregular interactions of centromere in early meiosis which may take participate in promoting meiosis stabilization in newly formed allotetraploid wheat. [ABSTRACT FROM AUTHOR]

Purpose: Asthenozoospermia is an important cause of male infertility, and the most serious type is characterized by multiple morphological abnormalities of the sperm flagella (MMAF). However, the precise etiology of MMAF remains unknown. In the current study, we recruited a consanguineous Pakistani family with two infertile brothers suffering from primary infertility due to MMAF without obvious signs of PCD. Methods: We performed whole-exome sequencing on DNAs of the patients, their parents, and a fertile brother and identified the homozygous missense variant (c.1490C > G (p.P497R) in NPHP4 as the candidate mutation for male infertility in this family. Results: Sanger sequencing confirmed that this mutation recessively co-segregated with the MMAF in this family. In silico analysis revealed that the mutation site is conserved across different species, and the identified mutation also causes abnormalities in the structure and hydrophobic interactions of the NPHP4 protein. Different bioinformatics tools predict that NPHP4p.P497R mutation is pathogenic. Furthermore, Papanicolaou staining and scanning electron microscopy of sperm revealed that affected individuals displayed typical MMAF phenotype with a high percentage of coiled, bent, short, absent, and/or irregular flagella. Transmission electron microscopy images of the patient's spermatozoa revealed significant anomalies in the sperm flagella with the absence of a central pair of microtubules (9 + 0) in every section scored. Conclusions: Taken together, these results show that the homozygous missense mutation in NPHP4 is associated with MMAF. [ABSTRACT FROM AUTHOR]

An algorithm based on the Paterson-Stockmeyer (PS) scheme and filtering technology is developed to compute the large matrix functions in dynamic problems accurately and efficiently. With the assistance of analysis on truncation error and error caused by filtering, the error growth law during the computation is studied, based on which an adaptive filtering threshold is proposed to help the proposed algorithm more efficiently achieve similar accuracy as the original PS scheme. Numerical examples, including 30 random matrices with different bandwidths, 10 adjacency matrices in the complex network dynamic problems, and a trampoline vibration problem, are given to verify the efficiency and accuracy of the proposed algorithm. Numerical results suggest that the proposed algorithm can achieve good accuracy and efficiency in computing the matrix function in the considered dynamic problems. [ABSTRACT FROM AUTHOR]

Key message: Complete new wheat-rye disomic, telosomic addition lines and various chromosomal aberrations were developed and characterized by molecular cytogenetic method as novel chromosome engineering materials. A new stem rust resistance (Ug99) gene was located on 3RL. Wheat stem rust, caused by Puccinia graminis f. sp. tritici (Pgt), is a devastating fungal disease worldwide. A recently emerged great threat to global wheat production is Pgt strain Ug99 and its derivatives, which have overcome most of the commonly used resistance genes. Rye (Secale cereale L.), closely related to wheat (Triticum aestivum L.), is a significant and valuable resource of resistance genes for wheat germplasm improvement. It is of great importance and urgency to identify new resistance gene sources of rye and transfer them into wheat. In this study, two complete sets of wheat-rye addition lines were established through wide hybridization, chromosome doubling and backcrossing. A wheat-rye 3RL telosomic addition line was identified with high resistance to stem rust strain Ug99. PCR-based markers specific for the rye chromosome were developed. Furthermore, abundant chromosomal aberrations such as minichromosomes, ring chromosomes as well as centromere reduction and expansion were identified in the progeny of wheat-rye addition lines by multicolor GISH and FISH. The line carrying a novel resistance gene to stem rust can be utilized as a bridge material for wheat disease resistance breeding. The chromosomal and centromeric variation within the wheat-rye hybrids can further contribute to genetic diversity of their offspring. [ABSTRACT FROM AUTHOR]

Background: Fascins belong to a family of actin-bundling proteins that are involved in a wide range of biological functions. FSCN3, a newly identified testis-specific actin-bundling protein, is specifically expressed in elongated spermatids. However, its in vivo function in mouse spermiogenesis remains unknown. Methods and results: We generated Fscn3 knockout mice through CRISPR/Cas9 gene-editing technology. Fscn3−/− mice displayed normal testis morphology and testis to bodyweight ratio, and sperm concentrations did not differ significantly between Fscn3+/+ and Fscn3−/− mice. Fertility assays consistently revealed that Fscn3−/− mice are completely fertile and their reproductive status does not differ from that of wild-type. Moreover, hematoxylin and eosin staining of the testis sections of Fscn3−/− mice detected various germ cells, ranging from spermatogonia to mature spermatozoa. Furthermore, the swimming velocity of the sperm of Fscn3−/− mice was comparable to that of their wild-type littermates. Both Fscn3+/+ and Fscn3−/−mice had normal sperm morphology, indicating that the disruption of Fscn3 does not affect sperm morphology. The analysis of meiotic prophase I progression demonstrated normal prophase-I phases (leptonema to diplonema) in both Fscn3+/+ and Fscn3−/− mice, suggesting that Fscn3 is not essential for meiosis I. Conclusion: Our study provides the first evidence that FSCN3 is a testis-specific actin-bundling protein that is not required for mouse spermatogenesis. Our results will help reproductive biologists focus their efforts on genes that are crucial for fertility and avoid research duplication. [ABSTRACT FROM AUTHOR]

Infertility is a multifactorial disorder that affects approximately 12% of couples of childbearing ages worldwide. Few studies have been conducted to understand the genetic causes of infertility in depth. The synaptonemal complex (SC), which is essential for the progression of meiosis, is a conserved tripartite structure that binds homologous chromosomes together and is thus required for fertility. This study investigated genetic causes of infertility in a Pakistani consanguineous family containing two patients suffering from non-obstructive azoospermia (NOA). We performed whole-exome sequencing, followed by Sanger sequencing, and identified a novel pathogenic variant (c.7G > A [p.D3N]) in the SC coding gene C14orf39, which was recessively co-segregated with NOA. In silico analysis revealed that charges on wild-type residues were lost, which may result in loss of interactions with other molecules and residues, and a reduction in protein stability occurred, which was caused by the p.D3N mutation. The novel variant generated the mutant protein C14ORF39D3N, and homozygous mutations in C14orf39 resulted in NOA. The transcriptome profile of C14ORF39 shows that it is specifically expressed in early brain development, which suggests that research in this area is required to study other functions of C14ORF39 in addition to its role in the germline. This research highlights the conserved role of C14orf39/SIX6OS1 in assembly of the SC and its indispensable role in facilitating genetic diagnosis in patients with infertility, which may enable the development of future treatments. [ABSTRACT FROM AUTHOR]

The WRKY family is a large group of transcription factors found in higher plants; it plays an important role in many aspects of biological processes. However, there is very little information about this family in melon (Cucumis melo L.). In our research, 57 candidate WRKY genes in the melon genome were identified. According to their structural and phylogenetic features, the 57 CmWRKY genes were classified into three groups, I, II, and III, and the group II was further divided into five subgroups. Group I included 11 members that all have two conservative WRKY domains and a C2H2-type zinc finger motif; Group II contains 41 WRKY gene family members, that all have a WRKY domain and a C2H2-type zinc finger motif. Five members that all have a conservative WRKY domain and a C2HC-type zinc finger motif are classified as Group III. The expression of 16 selected melon WRKY genes was detected by quantitative real-time PCR after sprayed with salicylic acid (SA) or powdery mildew infection. qRT-PCR analysis showed that 16 CmWRKY genes exhibited distinct expression patterns upon powdery mildew infection, and the expression levels of nine genes were inhibited, and seven genes were induced. After being sprayed with SA, the expression levels of 11 genes were inhibited, and five genes were induced. The data here provide an important basis for further functional studies of the WRKY gene in melon resistance. [ABSTRACT FROM AUTHOR]

Several genes have been reported to be involved in spermatogenesis but their functional importance in male fertility is yet needed to be elucidated. Therefore, in current research, we focused to explore the in vivo role of evolutionary conserved and testis-specifically expressed, C4orf46, gene in male mouse fertility and spermatogenesis. The expression profile of C4orf46 is specific to testes and expressed in testes from 7 days of postpartum to onward. Thus, we generated the C4orf46 knockout mice by utilizing CRISPR/Cas9 genome editing technology and examined gene function in spermatogenesis and fertility. Surprisingly, C4orf46 knockout mice were completely fertile, displayed normal testes morphology, however, higher sperm contents were observed in knockout mice compared to wild type (WT) littermates. Subsequently, intact testis histology and architecture of seminiferous tubules were observed in C4orf46 knockout and WT mice. Similarly, sperm morphology and swimming velocity of C4orf46 knockout mice were comparable with the WT littermates. Furthermore, all type of germ cells ranging from spermatogonia to mature spermatozoa were observed in the testes and epididymis sections of C4orf46 knockout mice suggesting that disruption of C4orf46 did not impact spermatogenesis. Moreover, meiotic prophase I progression was normal, and each type of cell population was comparable between knockout and WT mice. Overall, finding from this research indicates that C4orf46 is not an essential gene for fertility in mice. This study will help researchers to avoid the repetition and duplication of efforts, and to explore the genes that are indispensable for spermatogenesis and male fertility. [ABSTRACT FROM AUTHOR]

The photosynthesis, photomorphogenesis, and photoperiod processes in plants are regulated according to light intensity and quality. The aim of this study was to investigate the effects of different light qualities on eggplant seedlings and determine the best light quality for growth. The seedlings of eggplant cultivar 'Jingqiejingang' were grown under light-emitting diodes (LEDs): white (W, the control), red (R), blue (B), and different ratios of B/R lights (B/R = 1/1, B/R = 1/3, B/R = 1/6, B/R = 1/9). The growth parameters, leaf morphology, photosynthetic performance, chlorophyll fluorescence, and the carbon and nitrogen metabolism in the leaves of eggplant seedlings under different LED light treatments were studied. The results showed that the plant height, leaf development, and photosynthetic characteristics were inhibited by red light but elevated by blue light compared with the control. Conversely, the contents of chlorophyll a, chlorophyll b, and carotenoids were all increased by red light, while decreased by blue light significantly. In addition, the contents of carbohydrates and the activities of nitrogen assimilation enzymes were not or little changed by the monochromatic blue and red light. The combined light of red and blue were more beneficial for growth than the monochromatic light, especially B/R = 1/3 light. Under B/R = 1/3 light, the parameter values of plant growth, leaf development, photosynthetic pigments and characteristics, and carbon and nitrogen metabolism were all maximum. Taken together, combined application lights of red and blue are good practice for the cultivation of eggplant seedlings, and LED B/R = 1/3 light was optimum. [ABSTRACT FROM AUTHOR]

Background: Centromeres are specialized chromosomal domains involved in kinetochore formation and faithful chromosome segregation. Despite a high level of functional conservation, centromeres are not identified by DNA sequences, but by epigenetic means. Universally, centromeres are typically formed on highly repetitive DNA, which were previously considered to be silent. However, recent studies have shown that transcription occurs in this region, known as centromeric-derived RNAs (cenRNAs). CenRNAs that contribute to fundamental aspects of centromere function have been recently investigated in detail. However, the distribution, behavior and contributions of centromeric transcripts are still poorly understood. Objective: The aim of this article is to provide an overview of the roles of cenRNAs in centromere formation and function. Methods: We describe the structure and DNA sequence of centromere from yeast to human. In addition, we briefly introduce the roles of cenRNAs in centromere formation and function, kinetochore structure, accurate chromosome segregation, and pericentromeric heterochromatin assembly. Centromeric circular RNAs (circRNAs) and R-loops are rising stars in centromere function. CircRNAs have been successfully identified in various species with the assistance of high-throughput sequencing and novel computational approaches for non-polyadenylated RNA transcripts. Centromeric R-loops can be identified by the single-strand DNA ligation-based library preparation technique. But the molecular features and function of these centromeric R-loops and circRNAs are still being investigated. Conclusion: In this review, we summarize recent findings on the epigenetic regulation of cenRNAs across species, which would provide useful information about cenRNAs and interesting hints for further studies. [ABSTRACT FROM AUTHOR]

Although numerous studies have explored various impacts of Internet use, few have investigated the linkage between Internet use and chemical fertilizer use in developing economies. This study examines the influences of Internet use and related promotion policy on chemical fertilizer use based on a nationwide dataset including China's 7766 rural households. The baseline regression results show that Internet use reduced chemical fertilizer use significantly. The mechanism analysis unveils the mediation role of human capital; Internet use increased farmers' human capital and then reduced chemical fertilizer use. In addition, the difference-in-difference method is employed to examine the effects of Internet promotion policy and the findings reveal that the promotion policy could help reduce farmers' use of chemical fertilizer. Finally, both propensity score matching-DID model and a two-stage instrumental variable model are used to address potential endogeneity issues associated with Internet use, and results indicate that the empirical results are robust. Our findings suggest that improving the availability of broadband access in a rural area, promoting Internet-related training and education, and more government's investment in rural ICTs infrastructure can contribute to the reduction of chemical fertilizer use. [ABSTRACT FROM AUTHOR]

Reduced fertility is a common clinical feature of the individuals with Fanconi anemia (FA), a rare autosomal recessive disorder due to deficiency in FA pathway during DNA repair. Our previous study reported that the heterozygous pathogenic variants in FANCA (Fanconi anemia complementation group A) induced premature ovarian insufficiency (POI). However, the genotype–phenotype correlation in POI caused by FANCA variants remains considerably uncertain. Herein, a heterozygous non-frameshift Fanca-mutated mouse strain (Fanca+/hypo) carrying a 9-bp deletion (c.3581del9, p.QEA1194-1196del) was generated. The mutant mice exhibited slightly decreased Fanca protein level in ovaries, suggesting the non-frameshift deletion mutant is hypomorphic. Female fertility test showed decreased number of litters, litter sizes and prolonged litter interval time in the female Fanca+/hypo mice compared to wild-type mice. Follicle counting revealed a consistent decreasing pattern of follicle numbers in Fanca+/hypo females compared to that in wild-type mice with aging. Furthermore, embryonic fibroblasts of Fanca+/hypo mice were hyper-responsive to Mitomycin C in vitro, demonstrating a partial loss of function of this hypomorphic Fanca mutant in DNA repair. Collectively, our experimental observations suggest that the hypomorphic Fanca allele is sufficient to reduce female fertility in mice, providing new insights into the genetic counseling of FANCA variants in subfertile women. [ABSTRACT FROM AUTHOR]

Since the first discovery of old yellow enzyme 1 (OYE1) from Saccharomyces pastorianus in 1932, biocatalytic asymmetric reduction of activated alkenes by OYEs has become a valuable reaction in organic synthesis. To access stereocomplementary C=C-bond bioreduction, the mining of novel OYEs and especially the protein engineering of existing OYEs have been performed, which successfully achieved the stereocomplementary reduction in several cases and further raise the potential of applications. In this review, we analyzed the structures, active sites, and substrate recognition of OYEs, which are the bases for their substrate specificity and stereospecificity. Sequence similarity network of OYEs superfamily was also constructed to investigate the scope of characterized OYEs. The structure-guided engineering to switch the stereoselectivity of OYEs and thus access stereocomplementary bioreduction over the last decade (2009–2020) was then reviewed and discussed, which might give new insights into the mining and engineering of related biocatalysts. Key points: • The sequence similarity network of OYEs superfamily was constructed and annotated. • The structures and active sites of OYEs from different classes were compared. • "Left/right" binding mode was used to explain the stereopreferences of OYEs. • Structure-guided engineering of OYEs to switch their stereoselectivity was reviewed. [ABSTRACT FROM AUTHOR]

Mammalian mitochondria have small genomes encoding very limited numbers of proteins. Over one thousand proteins and noncoding RNAs encoded by the nuclear genome must be imported from the cytosol into the mitochondria. Here, we report the identification of hundreds of circular RNAs (mecciRNAs) encoded by the mitochondrial genome. We provide both in vitro and in vivo evidence to show that mecciRNAs facilitate the mitochondrial entry of nuclear-encoded proteins by serving as molecular chaperones in the folding of imported proteins. Known components involved in mitochondrial protein and RNA importation, such as TOM40 and PNPASE, interact with mecciRNAs and regulate protein entry. The expression of mecciRNAs is regulated, and these transcripts are critical for the adaption of mitochondria to physiological conditions and diseases such as stresses and cancers by modulating mitochondrial protein importation. mecciRNAs and their associated physiological roles add categories and functions to the known eukaryotic circular RNAs and shed novel light on the communication between mitochondria and the nucleus. [ABSTRACT FROM AUTHOR]

Melatonin is a natural phytohormone that occurs in most plants. It can regulate not only plant growth and development but also alleviate biotic and abiotic stress in plants. However, no studies have reported on endogenous melatonin increasing tomato tolerance to salt stress. In the present study, we observed that the overexpression of caffeic acid O-methyltransferase 1 (SlCOMT1) increased melatonin levels in tomato plants. We also observed that transgenic plants exhibited higher salt stress tolerance. SlCOMT1 overexpression could maintain the balance of Na+/K+ and decrease ion damage by activating salt overly sensitive (SOS) pathway under salt treatment. In addition, SlCOMT1 overexpression significantly enhanced the antioxidant capability, with higher antioxidant enzyme activity observed, including superoxide dismutase, peroxide and catalase activity, and higher ascorbic acid (AsA) and glutamate (GSH) accumulation levels. SlCOMT1 overexpression also maintained good nutrient homeostasis in the tomato plants. In addition, SlCOMT1 overexpression upregulated some stress-related genes (AREB1, AIM1, MAPK1, WRKY33 and CDPK1), which resulted in the activation of downstream signaling pathways and could be partly responsible for the improvement in salt stress tolerance. [ABSTRACT FROM AUTHOR]

Thousands of genes are involved in spermatogenesis, however, the functional roles of most these genes for male fertility remain to be discovered. This research focused to explore the function of evolutionarily conserved and testis-specific expressed gene 4930524B15Rik, which is known as C5orf47 in human. We generated 4930524B15Rik knockout mice by CRISPR/Cas9 technology and found 4930524B15Rik−/− mice were fertile. Furthermore, no averted abnormalities were observed in testis morphology, epididymal sperm contents and sperm morphology in 4930524B15Rik knockout mice. Subsequently, histological analysis of testicular tissue revealed intact structure of seminiferous tubules along with the presence of all types of germ cells in 4930524B15Rik−/− mice similar to wild type. Additionally, cytological analysis of spermatocytes displayed no significant differences in the prophase I progression of meiosis, further indicating that 4930524B15Rik have no essential function in mammalian spermatogenesis. Altogether, these results indicated that 4930524B15Rik is dispensable for fertility of male mice and these findings will help researchers to avoid future research overlap and to focus on genes that are crucial for spermatogenesis and reproduction. [ABSTRACT FROM AUTHOR]

Xanthine dehydrogenase (XDH) is an important enzyme in purine metabolism. It is involved in regulation of the normal growth and non-biological stress-induced ageing processes in plants. The present study investigated XDH's role in regulating rice leaf senescence. We measured physical characteristics, chlorophyll content and fluorescence parameters, active oxygen metabolism, and purine metabolism in wild-type Kitaake rice (Oryza sativa L.), an OsXDH over-expression transgenic line (OE9), and an OsXDH RNA interference line (Ri3) during different growth stages. The expression patterns of the OsXDH gene confirmed that XDH was involved in the regulation of normal and abiotic stress-induced ageing processes in rice. There was no significant difference between the phenotypes of transgenic lines and wild type at the seedling stage, but differences were observed at the full heading and maturation stages. The OE9 plants were taller, with higher chlorophyll content, and their photosystems had stronger light energy absorption, transmission, dissipation, and distribution capacity, which ultimately improved the seed setting rate and 1000-seed weight. The opposite effect was found in the Ri3 plants. The OE9 line had a strong ability to remove reactive oxygen species, with increased accumulation of allantoin and alantoate. Experimental spraying of allantoin on leaves showed that it could alleviate chlorophyll degradation and decrease the content of H2O2 and malonaldehyde (MDA) in rice leaves after the full heading stage. The urate oxidase gene (UO) expression levels in the interference line were significantly lower than those in the over-expression line and wild-type lines. The allantoinase (ALN) and allantoate amidinohydrolase (AAH) genes had significantly higher expression in the Ri3 plants than the in OE9 or wild-type plants, with OE9 plants showing the lowest levels. The senescence-related genes ACD1, WRKY23, WRKY53, SGR, XERO1, and GH27 in Ri3 plants had the highest expression levels, followed by those in the wild-type plants, with OE9 plants showing the lowest levels. These results suggest that enhanced activity of XDH can regulate the synthesis of urea-related substances, improve plant antioxidant capacity, effectively delay the ageing process in rice leaves, and increase rice yield. [ABSTRACT FROM AUTHOR]

Premature ovarian insufficiency (POI) is a major cause of reduced female fertility and affects approximately 1% women under 40 years of age. Recent advances emphasize the genetic heterogeneity of POI. Fanconi anemia (FA) genes, traditionally known for their essential roles in DNA repair and cytogenetic instability, have been demonstrated to be involved in meiosis and germ cell development. Here, we conducted whole-exome sequencing (WES) in 50 Han Chinese female patients with POI. Rare missense variants were identified in FANCA (Fanconi anemia complementation group A): c.1772G > A (p.R591Q) and c.3887A > G (p.E1296G). Both variants are heterozygous in the patients and very rare in the human population. In vitro functional studies further demonstrated that these two missense variants of FANCA exhibited reduced protein expression levels compared with the wild type, suggesting the partial loss of function. Moreover, mono-ubiquitination levels of FANCD2 upon mitomycin C stimulation were significantly reduced in cells overexpressing FANCA variants. Furthermore, a loss-of-function mutation of Fanca was generated in C57BL/6 mice for in vivo functional assay. Consistently, heterozygous mutated female mice (Fanca+/−) showed reduced fertility and declined numbers of follicles with aging when compared with the wild-type female mice. Collectively, our results suggest that heterozygous pathogenic variants in FANCA are implicated in non-syndromic POI in Han Chinese women, provide new insights into the molecular mechanisms of POI and highlight the contribution of FANCA variants in female subfertility. [ABSTRACT FROM AUTHOR]

The balanced actions between ubiquitination and deubiquitination precisely control the levels of various proteins vital for spermatogenesis. Ubiquitin-specific processing proteases (USPs) are the largest family of deubiquitinatingenzymes(DUBs), containing more than 50 members. So far, the functions of only a few USPs in male fertility have been studied, the roles of the majority are yet unknown. The present study aimed to explore the function of Usp29 (ubiquitin-specific protease 29) in male fertility. We found that Usp29 showed predominant expression in mouse testis, and its mRNA expression started to increase at 14 days postpartum (dpp), with a peak at 28 and 35 dpp. Using CRISPR/Cas9 technology, we generated Usp29 knockout mice (Usp29-/-). Usp29-/- mice exhibited no overt developmental anomalies. Further examination revealed that Usp29-/- mice had normal fertility and showed no detectable difference in the testis/body weight ratio, testicular and epididymal histology as well as epididymal sperm count from the wild-type littermates. Moreover, Usp29 is not a pseudogene in mice. Taken together, our study first reported that though Usp29 is predominantly expressed in the testis, it is not essential for male fertility in mice. [ABSTRACT FROM AUTHOR]

Aims: The present study was conducted to investigate the effectiveness of Streptomyces (GLY-P2) in degradation of ferulic acid (FA) and p-hydroxybenzonic acid (PHBA) in the rhizosphere of cucumbers by assaying the alteration of antioxidant enzymes activities and rhizospheric microbial community.Methods: GLY-P2 was isolated, identified as Streptomyces canus, and applied to cucumber-planted soil containing FA and PHBA.Results: Optimal conditions for FA and PHBA degradation by GLY-P2 were 40 °C, pH 7, and 0.2 g l−1 mixture of FA and PHBA. During the degradation, vanillin, vanillic acid, and protocatechoic acid were metabolites; and activities of superoxide dismutase (SOD), catalase, ascorbate peroxidase (APX), and dehydroascorbate reductase in GLY-P2 were induced. When inoculated into cucumber-planted soil containing 220 μg g−1 mixture of FA and PHBA, GLY-P2 degraded FA and PHBA in soil, improved plant growth, and decreased malonaldehyde, superoxide radical, and hydrogen peroxide levels in leaves. GLY-P2 also enhanced activities of SOD, catalase, glutathione peroxidase, APX, monodehydroascorbate reductase, dehydroascorbate reductase, and glutathione reductase, increased contents of ascorbate and glutathione, and elevated transcript levels of copper/zinc SOD, manganese SOD, catalase, and APX in leaves. Moreover, GLY-P2 changed soil bacterial richness, diversity, and community composition, and increased phosphatase, catalase, urease, and sucrase activities in rhizospheric soil.Conclusion: GLY-P2 mitigates FA and PHBA stress in cucumber by activating leaf antioxidant enzymes and affecting soil bacterial community. [ABSTRACT FROM AUTHOR]

We encourage centers with large cohorts of POI patients as well as FA patients to precisely define the genotype-phenotype correlations in women with I FANC i variants. To the Editor, We thank Heddar and Misrahi for their interest in our recent paper (Yang et al. [20]), and for sharing their different opinions on the causal link between I FANCA i variants and premature ovarian insufficiency (POI). First, no rare variant in any other causative/candidate POI genes has been found in our I FANCA i -associated patients after processing whole-exome sequencing (WES) data. [Extracted from the article]

A thermal deformation prediction model of a high-speed motorized spindle is important for improving machining accuracy and reducing the thermal error in the spindle. The convective heat transfer coefficient reflects the internal heat exchange capacity of a motorized spindle. In the finite element thermal analysis of a motorized spindle, the heat transfer coefficient is used as the boundary condition to calculate the temperature field and thermal deformation. The accuracy of the convective heat transfer coefficient has the most evident effect on the thermal deformation prediction of the motorized spindle. In this paper, a method for optimizing the heat transfer coefficient based on the biogeography optimization algorithm is proposed for the 100MD60Y4 motorized spindle. The proposed method is used to develop a thermal deformation prediction model of an intelligent accurate motorized spindle. Accurate prediction of the thermal deformation of the motorized spindle is realized using the experimental data of the surface temperature of the motorized spindle. Experimental results show that the average prediction error after the optimization of the spindle bearing temperature is 0.22 °C. The average prediction error in the thermal deformation of the spindle is 0.72 μm. The developed model is more accurate compared with the conventional thermal deformation prediction model of the motorized spindle. [ABSTRACT FROM AUTHOR]

Key message: Transcriptome data were used to develop 134 Aegilops longissima specific PCR markers and their comparative maps were constructed by contrasting with the homologous genes in the wheat B genome. Three wheat- Ae. longissima 1BL·1S S translocation lines were identified using the correspondence markers. Abstract: Aegilops longissima is an important wild species of common wheat that harbors many genes that can be used to improve various traits of common wheat ( Triticum aestivum L.). To efficiently transfer the traits conferred by these Ae. longissima genes into wheat, we sequenced the whole expression transcript of Ae. longissima. Using the transcriptome data, we developed 134 specific polymerase chain reaction markers located on the 14 chromosome arms of Ae. longissima. These novel molecular markers were assigned to specific chromosome locations based on a comparison with the homologous genes in the B genome of wheat. Annotation of these genes showed that most had functions related to metabolic processes, hydrolase activity, or catalytic activity. Additionally, we used these markers to identify three wheat- Ae. longissima 1BL·1SS translocation lines in somatic variation populations resulting from a cross between wheat cultivar Westonia and a wheat- Ae. longissima substitution line 1S(1B). The translocation lines had several low molecular weight glutenin subunits encoding genes beneficial to flour processing quality that came from Ae. longissima 1SS. The three translocation lines were also confirmed by genomic in situ hybridization. These translocation lines will be further evaluated for potential quality improvement of bread-making properties of wheat. [ABSTRACT FROM AUTHOR]

S- adenosylmethionine synthetase is a member of the stress-induced family genes. Our previous research indicated that overexpression of SlSAMS1 confers alkali stress tolerance to tomato seedlings. However, information regarding the alkali stress tolerance mechanism of SlSAMS1 and the cross-linked network between SlSAMS1 and downstream signal has been limited. To study how SlSAMS1 improves alkali stress tolerance, we manipulated the SlSAMS1 transgenic calluses through a pharmacological approach and found that overexpression of SlSAMS1 was positively correlated with polyamine (PA) and hydrogen peroxide (HO) accumulation leading to improve alkali stress tolerance. Additionally, the accumulation of HO in SlSAMS1 overexpression calluses depended on polyamine oxidase activity. The activities of antioxidant system, accumulation of organic acid, Na detoxification as well as alkali stress tolerance of the SlSAMS1 transgenic calluses were reversed by PA biosynthesis inhibitors, but not significantly influenced by ethylene biosynthesis inhibitors. These results suggest that overexpression of SlSAMS1 enhances alkali stress tolerance through PA and HO cross-linked networks, which provide new insight into how SlSAMS1 functions as a stress mediatory element in regulating plants tolerance to alkali stress. [ABSTRACT FROM AUTHOR]

Small supernumerary marker chromosomes (sSMCs) are structurally abnormal rare chromosomes, difficult to characterize by karyotyping, and have been associated with minor dysmorphic features, azoospermia, and recurrent miscarriages. However, sSMC with a gonosomal trisomy has never been reported. Spermatocyte spreading and immunostaining were applied to detect meiotic prophase I progression, homologous chromosome pairing, synapsis, and recombination. In all the analyzed spermatocytes of the patient, the extra Y chromosome was not detected while the sSMC was present. The recombination frequency on autosomes was not affected, while the recombination frequencies on XY chromosome was significantly lower in the patient than in the controls. The meiotic prophase I progression was disturbed with significantly increased proportion of zygotene and decreased pachytene spermatocytes in the patients as compared with the controls. These findings highlight the importance of studies on meiotic behaviors in patients with an abnormal chromosomal constitution and provide an important framework for future studies, which may elucidate the impairment caused by sSMC in mammalian meiosis and fertility. [ABSTRACT FROM AUTHOR]

Physiological responses of tomato roots to NaCl and NaHCO stresses were investigated in a hydroponic setting. The relative growth rate of tomato plants was significantly reduced in both NaCl and NaHCO treatments, especially under NaHCO stress. Tomato root respiration increased under low concentrations of NaCl and NaHCO stresses. However, high concentrations of both NaCl and NaHCO significantly inhibited respiration, especially in the NaHCO treatment. With increasing concentration of NaCl and NaHCO treatment, root Na accumulation increased, while accumulation of N, P, K, Fe, and Mg was significantly lower. Compared to NaCl, NaHCO treatment resulted in more dramatic changes in these nutrients. All organic acids investigated were increased by NaHCO after 5 days of treatment, but only oxalate, tartrate and malate were induced by NaCl. This implies that global regulation of organic acids might play an important role in tomato's alkali stress tolerance. Compared to NaCl treatments, NaHCO treatments induced much higher levels of reactive oxygen species (ROS) and lipid peroxidation after 5 days of treatment, which was accompanied by higher activities of antioxidant enzymes and higher concentrations of ascorbate-glutathione. However, after 10 days of treatment, 100 mM NaHCO stress led to lower accumulation of ROS, antioxidant enzyme activities, and ascorbate-glutathione content. This may have been because root metabolism had almost completely stopped, as indicated by lower root respiration and activity. [ABSTRACT FROM AUTHOR]

The objective of this study was to develop an efficient regeneration and transformation protocol for cucumber ( Cucumis sativus L.). Cotyledonary node explants showed a very high regeneration frequency (94.3 %) when incubated on Murashige and Skoog medium (MS) supplemented with 1.5 mg l 6-benzyladenine (BA). An Agrobacterium-mediated transformation methodology for cucumber was developed using this protocol. A construct containing a sense mitogen-activated protein kinase gene ( CsNMAPK) was used to transform cucumber cotyledonary nodes. Adventitious shoots were obtained in selection media containing 150 mg l kanamycin and 250 mg l carbenicillin 4 weeks after bacterial inoculation. A total of twelve transgenic plants were confirmed using PCR-specific analysis. The transgene copy number of four transgenic plants (random selection) was found to be a single gene copy using Southern blot analysis and the expression level of the CsNMAPK gene in the transgenic plants was significantly higher than in the wild-type plants. The transgenic rate was approximately 4.8 %. [ABSTRACT FROM AUTHOR]

The reproductive system of human female exhibits a much faster rate of aging than other body systems. Ovarian aging is thought to be dominated by a gradual decreasing numbers of follicles, coinciding with diminished quality of oocytes. Menopause is the final step in the process of ovarian aging. This review focuses on the mechanisms underlying the ovarian aging involving a poor complement of follicles at birth and a high rate of attrition each month, as well as the alternated endocrine factors. We also discuss the possible causative factors that contribute to ovarian aging, e.g., genetic factors, accumulation of irreparable damage of microenvironment, pathological effect and other factors. The appropriate and reliable methods to assess ovarian aging, such as quantification of follicles, endocrine measurement and genetic testing have also been discussed. Increased knowledge of the ovarian aging mechanisms may improve the prevention of premature ovarian failure. [ABSTRACT FROM AUTHOR]

A letter to the editor is presented that discusses a study on the behavior of lagging chromosomes and micronuclei in colorectal cancer cells.

Nitrate reductase is a key enzyme in the overall process of nitrate assimilation by plants. A full-length cDNA clone encoding nitrate reductase (NR; EC 1.6.6.1) was isolated from cucumber ( Cucumis sativus L.) by RT-PCR and RACE techniques. The NR of cucumber ( CsNR), a full-length cDNA sequence of 3032 bp contains an open reading frame of 2748 bp encoding 915 amino acids. The deduced 915 amino acid sequence showed high identities with NR from other plants. Quantitative real-time PCR analysis indicated that CsNR expression was different in root, stem, leaf, flower and mature fruit tissues. CsNR transcript level and nitrate reductase activity (NRA) was down-regulated and the change in NO concentration showed a negative trend with NRA in leaves when subjected to the 182 mM NO treatment. However, the CsNR transcript level was up-regulated in roots by 182 mM NO treatment. Furthermore, NRA in roots lagged behind CsNR expression and there was no obvious lag of NRA in leaves. This study found that in roots, there was no obvious relationship between NRA and NO content. These results indicated that NRA was not only controlled by the level of CsNR mRNA and there was an obvious negative relationship between NO content and NRA in leaves but not in roots. [ABSTRACT FROM AUTHOR]

Induced pluripotent stem (iPS) cells have greatly provoked people's interest due to their enormous potential of clinical applications. Increasing care is taken with the genetic safety of iPS cells. However, up to now, the chromosomal integrity of murine iPS (miPS) cells has been largely unknown. We have observed recurrent trisomy and/or Robertsonian translocation (Rb) of chromosome 14 in six out of nine independent miPS cell lines from three laboratories by G-banding, fluorescence in situ hybridization (FISH) and spectral karyotyping (SKY) analyses, while all the miPS cell lines were derived from mouse embryonic fibroblasts (MEFs) or neural precursor cells (NPCs) with a normal karyotype. The miPS cells with trisomy and/or Rb of chromosome 14 showed growth advantage over the miPS cells with a normal karyotype. We found a significantly higher frequency of Rbs in the miPS cell lines induced with c-Myc than those without c-Myc. Our findings demonstrate that miPS cell lines have the propensity for chromosomal aberrations and there is an obvious correlation between the extent of chromosomal aberrations in miPS cells and the transcriptional factors used for their reprogramming. Therefore, our study raises awareness of the need for improvements of the induction conditions of miPS cells in order to avoid the chromosomal aberrations and ensure future safe applications. [ABSTRACT FROM AUTHOR]

Paclitaxel is a widely used microtubule drug and cancer medicine. Here we report that by short exposure to paclitaxel at a low dose, multipolar spindles were induced in mitotic cells without centrosome amplification. Both TPX2 depletion and Aurora-A overexpression antagonized the multipolarity. Live cell imaging showed that some paclitaxel-treated cells accomplished multipolar cell division and a portion of the daughter cells went on to the next round of mitosis. The surviving cells grew into clones with varied genome content. The results indicated that an aneuploidy population could be induced by short exposure to paclitaxel at a low dose, implicating potential side effects of paclitaxel. [ABSTRACT FROM AUTHOR]

The effects of cow manure vermicompost on plant growth, metabolite contents, and antioxidant activities of Chinese cabbage were investigated in pot cultures. Five treatments were designed by mixing vermicompost and soil at ratio of 0:7, 1:7, 2:7, 4:7, 7:0 (w/w). Marketable weight of Chinese cabbage was significantly ( p < 0.05) higher in the 2:1 treatment than in the other treatments, while plants grown in the full soil treatment (0:7) showed the lowest marketable weight (Fig. 1a). Vermicompost application significantly increased the nutrient content of Chinese cabbage leaves ( p < 0.05), especially in the 4:7 treatment, with increases in the contents of soluble sugar (Fig. 2a), soluble protein (Fig. 2b), vitamin C (Fig. 3a), total phenols (Fig. 3b), and total flavonoids (Fig. 3c) by 62%, 18%, 200%, 25%, and 17% compared to the full soil treatment, respectively. The antioxidant activities expressed by 2, 2-Dipenyl-1-picrylhydrazyl-scavenging activity (Fig. 4a), hydroxyl (OH)-scavenging activity (Fig. 4b), and iron (Fe2+)-chelating activity (Fig. 4c) were higher by 92%, 40%, and 36% in the 4:7 than 0:7 the treatment, respectively. Vermicompost application significantly increased ( p < 0.05) the plant contents of 16 essential amino acids (Table 1); the total amino acid content showed the greatest increase in the 4:7 treatment, 90% compared to the full soil treatment. [ABSTRACT FROM AUTHOR]

Abstract  Actins are ubiquitous and highly conserved proteins that play key roles in cell formation and cellular activities. In this study, an actin gene was isolated from chickpea for the first time and designated as CarACT1 (for Cicer arietinum L. actin gene 1; Genbank accession no. EU529707). It encoded a putative protein with 377 amino acids and contained five exons and four introns within genomic DNA sequence. CarACT1 was localized in cytoplasm and showed high similarity to other well known actins from various species. Reverse transcription-polymerase chain reaction (RT-PCR) assay proved that CarACT1 transcripts were ubiquitously accumulated in all major organs, such as seedling roots, stems, leaves, flowers, young pods, and seeds, as well as in diverse developmental stages, such as leaf senescence, seed development and germination. Our results suggested that CarACT1 is an actin gene with physiological functions and may be served as a potential reference gene for transcription level of interesting genes in chickpea. [ABSTRACT FROM AUTHOR]

The plant-specific NAC (for NAM, ATAF1,2 and CUC2) proteins have been found to play important roles in plant development and stress responses. In this study, a NAC gene CarNAC1 (for Cicer arietinum L. NAC gene 1) was isolated from a cDNA library constructed with chickpea seedling leaves treated by polyethylene glycol. CarNAC1 encoded a putative protein with 239 amino acids and contained 3 exons and 2 introns within genomic DNA sequence. CarNAC1 had a conserved NAC domain in the N-terminus and the CarNAC1:GFP (green fluorescent protein) fusion protein was localized in the nucleus of onion epidermal cells. Additionally, CarNAC1 exhibited the trans-activation activity which was mapped to the C-terminus. The CarNAC1 transcript was detected in many chickpea organs including seedling leaves, stems, roots, flowers, and young pods, but less accumulated in young seeds. CarNAC1 was induced by leaf age and showed changes in expression during seed development and germination. Furthermore, the expression of CarNAC1 was strongly induced by drought, salt, cold, wounding, H2O2, ethephon, salicylic acid, indole-3-acetic acid, and gibberellin. Our results suggest that CarNAC1 encodes a novel NAC-domain protein and may be a transcriptional activator involved in plant development and various stress responses. [ABSTRACT FROM AUTHOR]

Collaborator of ARF (CARF) regulates cell proliferative fate through both p53-dependent and -independent mechanisms. Recently, we reported a new function of CARF as a positive regulator of Wnt signaling. Despite these findings, the physiological function of CARF has not been well studied. Here, we generated CARF knockout mice and found that male CARF−/− mice exhibited significantly impaired fertility and Sertoli-cell-only (SCO) syndrome phenotypes. Further studies revealed that loss of CARF in Sertoli cells led to decreased GDNF expression, which hindered spermatogonial stem cells (SSCs) self-renewal. Meanwhile, CARF loss in undifferentiated spermatogonia impaired their proliferation. These two mechanisms together led to SCO syndrome phenotypes, which could be functionally rescued by pharmacological or genetic reactivation of Wnt signaling. Finally, we identified CARFS351F as a potential pathogenic mutation in an SCO patient. Overall, our findings reveal important roles of CARF in spermatogonial self-renewal and proliferation through the Wnt signaling pathway. [ABSTRACT FROM AUTHOR]

Males with a 47,XYY karyotype generally have chromosomally normal children, despite the high theoretical risk of aneuploidy. Studies of sperm karyotypes or FISH analysis of sperm have demonstrated that the majority of sperm are chromosomally normal in 47,XYY men. There have been a number of meiotic studies of XYY males attempting to determine whether the additional Y chromosome is eliminated during spermatogenesis, with conflicting results regarding the pairing of the sex chromosomes and the presence of an additional Y. We analyzed recombination in the pseudoautosomal region of the XY bivalent to determine whether this is perturbed in a 47,XYY male. A recombination frequency similar to normal 46,XY men would indicate normal pairing within the XY bivalent, whereas a significantly altered frequency would suggest other types of pairing such as a YY bivalent or an XYY trivalent. Two DNA markers, STS/STS pseudogene and DXYS15, were typed in sperm from a heterozygous 47,XYY male. Individual sperm (23,X or Y) were isolated into PCR tubes using a FACStarPlus flow cytometer. Hemi-nested PCR analysis of the two DNA markers was performed to determine the frequency of recombination. A total of 108 sperm was typed with a 38% recombination frequency between the two DNA markers. This is very similar to the frequency of 38.3% that we have observed in 329 sperm from a normal 46,XY male. Thus our results suggest that XY pairing and recombination occur normally in this 47,XYY male. This could occur by the production of an XY bivalent and Y univalent (which is then lost in most cells) or by loss of the additional Y chromosome in some primitive germ cells or spermatogonia and a proliferative advantage of the normal XY cells. [ABSTRACT FROM AUTHOR]

The N-myc downstream regulated gene (NDRG) family consists of 4 members, NDRG-1, -2, -3, -4. Physiologically, we found Ndrg3, a critical gene which led to homologous lethality in the early embryo development, regulated the male meiosis in mouse. The expression of Ndrg3 was enhanced specifically in germ cells, and reached its peak level in the pachytene stage spermatocyte. Haplo-insufficiency of Ndrg3 gene led to sub-infertility during the male early maturation. In the Ndrg3+/− germ cells, some meiosis events such as DSB repair and synaptonemal complex formation were impaired. Disturbances on meiotic prophase progression and spermatogenesis were observed. In mechanism, the attenuation of pERK1/2 signaling was detected in the heterozygous testis. With our primary spermatocyte culture system, we found that lactate promoted DSB repair via ERK1/2 signaling in the male mouse germ cells in vitro. Deficiency of Ndrg3 gene attenuated the activation of ERK which further led to the aberrancy of DSB repair in the male germ cells in mouse. Taken together, we reported that Ndrg3 gene modulated the lactate induced ERK pathway to facilitate DSB repair in male germ cells, which further regulated meiosis and subsequently fertility in male mouse. [ABSTRACT FROM AUTHOR]

ECAT1 is a subunit of the subcortical maternal complex that is required for cell cycle progression during pre-implantation embryonic development; however, its exact function remains to be elucidated. Here we investigated the expression of ECAT1 in human ovarian tissue, oocytes and pre-implantation embryos and assessed its function by using RNA interference (RNAi) in oocytes. ECAT1 mRNA was highly expressed in human oocytes and zygotes, as well as in two-cell, four-cell and eight-cell embryos, but declined significantly in morulae and blastocysts. ECAT1 was expressed in the cytoplasm of oocytes and pre-implantation embryos and was localized more specifically in the cortical region than in the inner cytoplasm. RNAi experiments demonstrated that down-regulation of ECAT1 expression not only impaired spindle assembly and reduced maturation and fertilization rates of human oocytes but also decreased the cleavage rate of the resulting zygotes. In conclusion, our study indicates that ECAT1 may play a role in meiotic progression by maintaining the accuracy of spindle assembly in human oocytes, thus promoting oocyte maturation and subsequent development of the embryo. [ABSTRACT FROM AUTHOR]

Weaver et al. report that chromosome nondisjunction induced by knockout of the mitosis-specific motor protein CENP-E does not increase the rate of binucleation in mouse fibroblasts and conclude that nondisjunction is therefore not coupled to furrow regression. However, they have not yet excluded alternative interpretations of their results, which makes it hard to compare them directly to our findings. Critical confounding issues include analysis of different cell types and the use of a specific gene deletion, for which corresponding naturally occurring mutations have not been described. [ABSTRACT FROM AUTHOR]