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Plant mineral nutrition is critical for agricultural productivity and for human nutrition; however, the availability of mineral elements is spatially and temporally heterogeneous in many ecosystems and agricultural landscapes. Nutrient imbalances trigger intricate signalling networks that modulate plant acclimation responses. One signalling agent of particular importance in such networks is phytomelatonin, a pleiotropic molecule with multiple functions. Evidence indicates that deficiencies or excesses of nutrients generally increase phytomelatonin levels in certain tissues, and it is increasingly thought to participate in the regulation of plant mineral nutrition. Alterations in endogenous phytomelatonin levels can protect plants from oxidative stress, influence root architecture, and influence nutrient uptake and efficiency of use through transcriptional and post-transcriptional regulation; such changes optimize mineral nutrient acquisition and ion homeostasis inside plant cells and thereby help to promote growth. This review summarizes current knowledge on the regulation of plant mineral nutrition by melatonin and highlights how endogenous phytomelatonin alters plant responses to specific mineral elements. In addition, we comprehensively discuss how melatonin influences uptake and transport under conditions of nutrient shortage.

Background: Male infertility is a major health concern in couples of childbearing ages. Nonobstructive azoospermia (NOA) is an extreme form of male infertility that affects ∼1% of adult men, and th...

Melatonin (MT) is a phytohormone-like substance and is profoundly involved in modulating nearly all aspects of plant development and acclimation to environmental stressors. However, there remain no studies about the effects of MT on tomato seed germination under salt stress. Here we reported that the overexpression of

Following meiotic recombination, each pair of homologous chromosomes acquires at least one crossover, which ensures accurate chromosome segregation and allows reciprocal exchange of genetic information. Recombination failure often leads to meiotic arrest, impairing fertility, but the molecular basis of recombination remains elusive. Here, we report a homozygous M1AP splicing mutation (c.1074 + 2T C) in patients with severe oligozoospermia owing to meiotic metaphase I arrest. The mutation abolishes M1AP foci on the chromosome axes, resulting in decreased recombination intermediates and crossovers in male mouse models. M1AP interacts with the mammalian ZZS (an acronym for yeast proteins Zip2-Zip4-Spo16) complex components, SHOC1, TEX11, and SPO16. M1AP localizes to chromosomal axes in a SPO16-dependent manner and colocalizes with TEX11. Ablation of M1AP does not alter SHOC1 localization but reduces the recruitment of TEX11 to recombination intermediates. M1AP shows cytoplasmic localization in fetal oocytes and is dispensable for fertility and crossover formation in female mice. Our study provides the first evidence that M1AP acts as a copartner of the ZZS complex to promote crossover formation and meiotic progression in males.

Polyploidy is a common mode of evolution in flowering plants. Both the natural tetraploid Thinopyrum elongatum and the diploid one from the same population show a diploid-like pairing in meiosis. However, debate on the chromosome composition and origin of the tetraploid Th. elongatum is ongoing. In the present study, we obtained the induced tetraploid Th. elongatum and found that the induced and natural tetraploids are morphologically close, except for slower development and lower seed setting. Using probes developed from single chromosome microdissection and a Fosmid library, obvious differentiations were discovered between two chromosome sets (E

PurposeChina is the world's largest consumer of pesticides. To increase the use efficiency and achieve more sustainable and environmentally friendly use of pesticides in China, it is crucial to understand why Chinese farmers use such a large amount of pesticides.Design/methodology/approachThe relationship between farm size and pesticide use was investigated by using national household-level panel data from 1995 to 2016.FindingFarms that are small and fragmented lead to the use of large amounts of pesticides in China. For a given crop type, three factors contribute to a negative relationship between farm size and pesticide use: the spillover effect from the use of pesticides by other farmers in the same village, the level of mechanization and the management ability of farmers. The first two factors play important roles in the cultivation of grain crops, while the last factor is the main reason why farmers with larger plots of land use fewer pesticides in the cultivation of vegetables. In addition, the effect of agricultural machinery services on reducing the use of pesticides is currently limited, and the service system in China is still insufficient, which has been pointed out that it is also due to the prevalence of small and fragmented farms.Originality/valueThe authors investigate and compare the farm size–pesticide use relationship in both grain and cash crop production. Moreover, the authors systematically explore and explain how farm size is related to a reduction in pesticide use in the cultivation of grain crops and cash crops. These results can help to better understand the role of land scale in pesticide use, lay a foundation for the formulation of policies to reduce pesticide use and provide valuable knowledge about pesticide use for other developing countries around the world.

Family with sequence similarity 181 (Fam181) is a gene family with two paralogues (Fam181a and Fam181b) found among vertebrates. Fam181a exhibits dynamic and stage-specific expression during murine embryo development. Furthermore, searching in the National Center for Biotechnology Information database revealed predominant expression of Fam181a in mouse and human testes, implying that it may have essential roles in spermatogenesis. In this study we investigated the in vivo function of Fam181a in mouse spermatogenesis and fertility by generating Fam181a–/– mice using clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) 9 genome editing technology. The resulting Fam181a–/– mice exhibited normal growth and development. In addition, the mice were completely fertile, with no obvious differences in the testis-to-bodyweight ratio, epididymal sperm count or sperm motility compared with wild-type mice. Further examination of testicular and epididymal histology of Fam181a–/– mice found an intact seminiferous tubule structure and the presence of all types of germ cells, from spermatogonia to mature spermatozoa, similar to wild-type littermates. Similarly, analysis of meiotic prophase I progression revealed normal populations of each substage of prophase I in Fam181a+/+ and Fam181a–/– testes, suggesting that this gene is dispensable for male fertility. These negative findings will help avoid research overlap, save time and resources and allow researchers to concentrate on genes that are critical for male fertility and spermatogenesis.

Non-obstructive azoospermia (NOA) is a severe factor of male infertility; it affects approximately 1% of the global male population and accounts for 40% of male infertility cases. However, the majority of NOA cases remain idiopathic. This is the first study using whole-exome sequencing (WES) to identify a novel missense mutation in the DND1 gene (c.212A>C, p. E71A) from a Pakistani family, that includes three males with NOA. This mutation is predicted to cause DND1 protein misfolding and weaken the DND1 interaction with NANOS2, a significant regulator in primordial germ cell development. Our study identified a DND1 pathogenic mutation in NOA patients and highlighted its critical role in male fertility in humans.

Testis-specifically expressed genes are important for male reproduction according to their unique expression patterns. However, the functions of most of these genes in reproduction are unclear. Here, we showed that mouse 4930590J08Rik was a testis-specifically expressed gene. 4930590J08Rik knockout mice exhibited a delay in the first wave of spermatogenesis and a reduction of cauda epididymal sperm. Furthermore, knockout spermatozoa exhibited defective acrosome reactions and decreased progressive motility, which led to impaired in vivo fertilization. Transcriptome analysis of testes revealed that most of the differentially expressed genes in knockout testes were associated with metabolic processes. 4930590J08Rik knockout sperm exhibited oxidative phosphorylation deficiency and were highly dependent on increased anaerobic glycolysis to compensate for ATP demands. Taken together, the 4930590J08Rik-disrupted mouse partially mimics the phenotypes of human asthenospermia and oligozoospermia, which provides a new model for further understanding the pathogenesis of idiopathic male infertility.

Sperm-oocyte membrane fusion is necessary for mammalian fertilization. The factors that determine the fusion of sperm with oocytes are largely unknown. So far, spermatozoon factor IZUMO1 and the IZUMO1 counter-receptor JUNO on the oocyte membrane has been identified as a protein requiring fusion. Some sperm membrane proteins such as FIMP, SPACA6 and TEME95, have been proved not to directly regulate fusion, but their knockout will affect the fusion process of sperm and oocytes. Here, we identified a novel gene C11orf94 encoding a testicular-specific small transmembrane protein that emerges in vertebrates likely acquired via horizontal gene transfer from bacteria and plays an indispensable role in sperm-oocyte binding. We demonstrated that the deletion of C11orf94 dramatically decreased male fertility in mice. Sperm from C11orf94-deficient mice could pass through the zona pellucida, but failed to bind to the oocyte membrane, thus accumulating in the perivitelline space. In consistence, when the sperm of C11orf94-deficient mice were microinjected into the oocyte cytoplasm, fertilized oocytes were obtained and developed normally to blastocysts. Proteomics analysis revealed that C11orf94 influenced the expression of multiple gene products known to be indispensable for sperm-oocyte binding and fusion, including IZUMO1, EQTN and CRISP1. Thus, our study indicated that C11ORF94 is a vertebrate- and testis-specific small transmembrane protein that plays a critical role in sperm binding to the oolemma.

During meiosis, at least one crossover must occur in each chromosome pair to ensure their accurate segregation. However, the mechanism of crossover formation is poorly understood. Here, we report a previously uncharacterized recombination protein, C12ORF40/REDIC1, essential for meiotic crossover formation. A homozygous frameshift mutation in C12orf40 was identified in two genetically unrelated infertile men with meiotic arrest. Subsequent analysis showed that REDIC1 forms discrete foci in the paired regions of homologous chromosomes depending on strand invasion. Mouse models mimicking the patients’ mutation showed reductions in crossover and bivalent formation, leading to spermatogenic failure. Notably, mutant spermatocytes displayed reduced numbers of MSH4 and TEX11 foci, accompanied by synaptic defects. Correspondingly, these phenotypes were also observed in our patients. Finally, biochemical results showed that the identified mutation impairs its ability to bind branched recombination intermediates. Therefore, our findings reveal a crucial role for C12ORF40/REDIC1 in meiotic crossover formation by stabilizing the recombination intermediates, providing prospective molecular targets for the clinical diagnosis and therapy of infertility.

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.

STUDY QUESTION Are mutations in the zinc finger SWIM domain-containing protein 7 gene (ZSWIM7) associated with human male infertility? SUMMARY ANSWER The homozygous frameshift mutation (c.231_232del) in ZSWIM7 causes decreased meiotic recombination, spermatogenesis arrest, and infertility in men. WHAT IS KNOWN ALREADY ZSWIM7 is a SWIM domain-containing Shu2/SWS1 protein family member and a subunit of the Shu complex. Zswim7 knockout mice were infertile due to impaired meiotic recombination. However, so far there is no direct evidence that mutations of ZSWIM7 cause human infertility. STUDY DESIGN, SIZE, DURATION Screening for mutations of ZSWIM7 was performed using in-house whole-exome sequencing data from 60 men with non-obstructive azoospermia (NOA). Mice with a corresponding Zswim7 mutation were generated for functional verification. PARTICIPANTS/MATERIALS, SETTING, METHODS Sixty Chinese patients, who were from different regions of China, were enrolled. All the patients were diagnosed with NOA owing to spermatocyte maturation arrest based on histopathological analyses and/or immunostaining of spermatocyte chromosome spreads. ZSWIM7 mutations were screened from the whole-exome sequencing data of these patients, followed by functional verification in mice. MAIN RESULTS AND THE ROLE OF CHANCE A homozygous frameshift mutation (c.231_232del) in ZSWIM7 was found in two out of the 60 unrelated NOA patients. Both patients displayed small testicular size and spermatocyte maturation arrest in testis histology. Spermatocyte chromosome spreads of one patient revealed meiotic maturation arrest in a pachytene-like stage, with incomplete synapsis and decreased meiotic recombination. Male mice carrying a homozygous mutation similar to that of our patients were generated and also displayed reduced recombination, meiotic arrest and azoospermia, paralleling the spermatogenesis defects in our patients. LIMITATIONS, REASONS FOR CAUTION As Zswim7 is also essential for meiosis in female mice, future studies should evaluate the ZSWIM7 mutations more in depth and in larger cohorts of infertile patients, including males and females, to validate the findings. WIDER IMPLICATIONS OF THE FINDINGS These findings provide direct clinical and functional evidence that the recurrent ZSWIM7 mutation (c.231_232del) causes decreased meiotic recombination and leads to male infertility, illustrating the genotype-phenotype correlations of meiotic recombination defects in humans. STUDY FUNDING/COMPETING INTEREST(S) This work was supported by the National Natural Science Foundation of China (31890780, 31630050, 32061143006, 82071709, and 31871514), the Strategic Priority Research Program of the Chinese Academy of Sciences (XDB19000000), and the National Key Research and Developmental Program of China (2018YFC1003900 and 2019YFA0802600). TRIAL REGISTRATION NUMBER Not applicable.

Asthenoteratozoospermia characterized by multiple morphological abnormalities of the flagella (MMAF) has been identified as a sub-type of male infertility. Recent progress has identified several MMAF-associated genes with an autosomal recessive inheritance in human affected individuals, but the etiology in approximately 40% of affected individuals remains unknown. Here, we conducted whole-exome sequencing (WES) and identified hemizygous missense variants in the X-linked CFAP47 in three unrelated Chinese individuals with MMAF. These three CFAP47 variants were absent in human control population genome databases and were predicted to be deleterious by multiple bioinformatic tools. CFAP47 encodes a cilia- and flagella-associated protein that is highly expressed in testis. Immunoblotting and immunofluorescence assays revealed obviously reduced levels of CFAP47 in spermatozoa from all three men harboring deleterious missense variants of CFAP47. Furthermore, WES data from an additional cohort of severe asthenoteratozoospermic men originating from Australia permitted the identification of a hemizygous Xp21.1 deletion removing the entire CFAP47 gene. All men harboring hemizygous CFAP47 variants displayed typical MMAF phenotypes. We also generated a Cfap47-mutated mouse model, the adult males of which were sterile and presented with reduced sperm motility and abnormal flagellar morphology and movement. However, fertility could be rescued by the use of intra-cytoplasmic sperm injections (ICSIs). Altogether, our experimental observations in humans and mice demonstrate that hemizygous mutations in CFAP47 can induce X-linked MMAF and asthenoteratozoospermia, for which good ICSI prognosis is suggested. These findings will provide important guidance for genetic counseling and assisted reproduction treatments.

PurposeThe purpose of this paper is to investigate how China's rural public pension affects farmers' formal borrowing, which has always been rationed.Design/methodology/approachThis paper uses a difference-in-difference (DID) estimation to evaluate the effect of the implementation of the New Rural Pension Scheme (NRPS) at the end of 2009 on farmers' formal borrowing.FindingsThe results show that the NRPS significantly reduces farmers' formal borrowing from rural credit cooperatives (RCCs). The effect is significant among the elderly, eastern China and high-income groups.Originality/valueThis study contributes to the literature by identifying another potential reason for rural formal credit shortage. Policymakers and rural formal financial institutions should consider the demand side problem of lending.

Long-term straw return is an important carbon source for improving soil organic carbon (SOC) stocks in croplands, and straw removal through burning is also a common practice in open fields in South China. However, the specific effects of long-term rice straw management on SOC fractions, the related enzyme activities and their relationships, and whether these effects differ between crop growing seasons remain unknown. Three treatments with equal nitrogen, phosphorus, and potassium nutrient inputs, including straw/ash and chemical nutrients, were established to compare the effects of straw removal (CK), straw return (SR), and straw burned return (SBR). Compared to CK, long-term SR tended to improve the yield of early season rice (P=0.057), and significantly increased total organic carbon (TOC) and microbial biomass carbon (MBC) in double-cropped rice paddies. While SBR had no effect on TOC, it decreased light fraction organic carbon (LFOC) in early rice and easily oxidizable organic carbon (EOC) in late rice, significantly increased dissolved organic carbon (DOC), and significantly decreased soil pH. These results showed that MBC was the most sensitive indicator for assessing changes of SOC in the double-cropped rice system due to long-term straw return. In addition, the different effects on SOC fraction sizes between SR and SBR were attributed to the divergent trends in most of the soil enzyme activities in the early and late rice that mainly altered DOC, while DOC was positively affected by β-xylosidase in both early and late rice. We concluded that straw return was superior to straw burned return for improving SOC fractions, but the negative effects on soil enzyme activities in late rice require further research.

Biofumigation is an environmentally friendly strategy used to control nematodes and plant diseases. The volatile oil of Zanthoxylum bungeanum has high insecticidal and antibacterial activity. However, it is not known if the seed of Z. bungeanum is a suitable material for biofumigation to control southern root-knot nematodes (SRKN) on tomato, and how it may regulate the soil bacterial community structure. We used pot experiments in the greenhouse to determine the effects of Z. bungeanum seeds on SRKN, plant growth parameters, soil physicochemical and microbial characteristics. A total of 26 volatile components, including nematicidally active substances, were identified from Z. bungeanum seeds. Z. bungeanum seed biofumigation significantly reduced the SRKN population by 88.89% and 81.55% on the 50th and 100th day after transplanting, respectively. Compared to the control, the total soluble sugar, soluble solids, soluble protein, titratable acid, root activity and the fruit yield per tomato plant increased significantly. The content of soil alkali-hydrolyzed nitrogen, available potassium and the soil enzyme activities were also significantly increased. The soil bacterial diversity and the co-occurrence network complexity were increased by Z. bungeanum seed biofumigation. Relatively more keystone OTUs in biofumigation soil had potential plant growth-promoting capabilities. The function of Z. bungeanum seed increasing tomato production in SRKN-infected soil depends on directly killing SRKN and improving soil properties. These results indicate that Z. bungeanum seed can be used as both a nematicide and a high quality organic fertilizer in tomato production.

Non-obstructive azoospermia (NOA), characterized by spermatogenesis failure and the absence of sperm in ejaculation, is the most severe form of male infertility. However, the etiology and pathology between meiosis-associated monogenic alterations and human NOA remain largely unknown. A homozygous

Meiosis is pivotal for sexual reproduction and fertility. Meiotic programmed DNA double-strand breaks (DSBs) initiate homologous recombination, ensuring faithful chromosome segregation and generation of gametes. However, few studies have focused on meiotic DSB formation in human reproduction. Here, we report four infertile siblings born to a consanguineous marriage, with three brothers suffering from non-obstructive azoospermia and one sister suffering from unexplained infertility with normal menstrual cycles and normal ovary sizes with follicular activity. An autosomal recessive mutation in TOP6BL was found co-segregating with infertility in this family. Investigation of one male patient revealed failure in programmed meiotic DSB formation and meiotic arrest prior to pachytene stage of prophase I. Mouse models carrying similar mutations to that in patients recapitulated the spermatogenic abnormalities of the patient. Pathogenicity of the mutation in the female patient was supported by observations in mice that meiotic programmed DSBs failed to form in mutant oocytes and oocyte maturation failure due to absence of meiotic recombination. Our study thus illustrates the phenotypical characteristics and the genotype-phenotype correlations of meiotic DSB formation failure in humans.

The ubiquitin proteins play important role in proteasomal degradation and their balanced action is essential for the crucial process of spermatogenesis. The disruption of various ubiquitinating proteins in mice revealed defective spermatogenesis, thus inferring their important function in spermatogenesis. However, the role of some testis-specific ubiquitinating proteins still needs to be discovered. This study was planned to study the in vivo function of testis-specific and evolutionarily conserved ubiquitin shuttle gene, Ddi1 (DNA damage inducible 1). Ddi1 knockout mice were generated by CRISPR/Cas9 technology and we found that Ddi1 knockout mice were fertile without obvious alterations in reproductive parameters, such as sperm number and morphology. Histological examination of testicular tissues manifested compact seminiferous tubule structure along with all type of germ cells in the knockout mice. Moreover, cytological studies of spermatocytes did not exhibit any noteworthy difference in the progression of prophase I which endorse the fact that Ddi1 has not any vital function during meiosis. Overall, these findings suggested that Ddi1 is not critical for mouse fertility under normal laboratory conditions. The outcome of this study will help researchers to avoid overlap that will not only save their resources but also concentrate their focus on indispensable genes in spermatogenesis and fertility.

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.

Plant development and structure are constantly adjusted throughout their life cycle to adapt environmental changes in nature. The amount of branches mainly depends on whether axillary buds form, release and growth of axillary buds. The plants branching is regulated by many factors including gene transcription, hormone homeostasis, and environmental factors such as light, water, and nutrition. People control shoot branching according to their demands to facilitate agricultural production. The activation and growth of lateral branches depend on a variety of internal and external environmental signals. Here we reviewed the recent progresses on genes, hormones, signaling materials, and external environmental conditions on the development of lateral branches in plant. Understanding these key nodes or regulatory networks is necessary and worthy challenges. The discussion of the factors affecting the formation of lateral branches provides a basis for the rational method of crop breeding and the cultivation of ideal plant type crops.

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.

To better understand cytokinin signaling in melon (Cucumis melo L.), one of the most important fruit crops in the Cucurbitaceae family, we identified and characterized melon two-component system (TCS) genes in this study. The results showed that there were 51 genes encoding putative TCS proteins in melon, and these TCS genes were classified into 3 subgroups, with 17 HK(L)s (histidine kinase/histidine-kinase like; 9 HKs and 8 HKLs), 9 HPs (histidine phosphotransfer proteins; 6 authentic and 3 pseudo), and 25 RRs (response regulators; 8 Type-A, 11 Type-B and 6 pseudo). The identity values of these cytokinin signaling proteins were revealed by analyzing their conserved motifs, domains and amino acid sequences. By analyzing TCS genes in different plant species, we found that melon HK(L)s, HPs and RRs had closer phylogenetic relationships with cucumber genes than with the genes of other plants, and the expansion of melon cytokinin signaling genes might be attributed to segmental duplication events. Analysis of the putative promoter regions (2-kb upstream regions of the start codon) revealed the enrichment of stress- and hormone-response cis-elements. The involvement of these putative TCS genes in melon cytokinin signaling was further supported by qRT-PCR data.

Lateral branches (LBs) pruning is performed frequently to keep the tomato plants in optimal growth conditions. However, the suitable pruning length of LBs, as well as the physiological and molecular mechanisms of pruning length on plant growth regulation remains elusive in tomato. The effects of pruning length of LBs from 0 to 20 cm on vegetative growth, reproductive growth, labor costs, hormone metabolism and genes transcripts were evaluated in an indeterminate type tomato cultivar. By comprehensive analysis, we provided evidence that pruning length of LBs at about 6–7 cm was suitable for plant growth, high yield, and low labor costs in tomato production. For mechanisms, appropriate extension of pruning length of LBs increased indole acetic acid (IAA) concentrations in root, which promoted the biosynthesis and upward transport of inactive cytokinins (CKs), as well as root development. Meanwhile, existence of LBs inhibited the auxin outflow of the lower flower stalks by regulating transcripts of AUX1 and PIN. Then, the enhanced concentrations of IAA and CKs in ovary could promote fruit swelling. Additionally, pruning length of LBs also influenced the leaf senescence to control photosynthesis. Taken together, we highlight that pruning length of LBs influence auxin and cytokinins homeostasis in relation to growth and yield in tomato plants.

Hierarchical nano-ZSM-5 zeolites (Z5-X) with different grain sizes were synthesized by varying amounts of 3-glycidoxypropyltrimethoxysilane (KH-560) in the hydrothermal synthesis strategy. Moreover, the conventional ZSM-5zeolite(Z5), which was prepared without KH-560, was used as the reference sample. The crystalline phases, morphologies, porous characteristics, Si/Al molar ratios and acidic properties of all fresh catalysts were characterized using the X-ray diffraction (XRD), Fourier transform infrared spectra (FT-IR), scanning electron microscopy (SEM), N2 adsorption–desorption, inductively coupled plasma atomic emission spectroscopy (ICP) and temperature programmed desorption of ammonia (NH3-TPD) techniques. Results show that the grain size and strong acid amount of zeolite decreased with the increasing amount of KH-560. The micropore surface areas and the corresponding volume of Z5-X changed less compared with Z5. Consequently, the high shape-selectivity of zeolite was preserved well under the addition of KH-560. However, the mesopore surface areas and the corresponding volume increased significantly with the increasing amount of KH-560. Benefiting from the abundant hierarchical structure, the Z5-X catalysts exhibited a larger coke capacity than the Z5 catalyst. The coke depositions of all the deactivated catalysts were characterized by the thermogravimetric technique (TG), and the results are indicative of the decreased average rate of coke deposition with an increasing amount of KH-560, which could result from the gradually reduced strong acid amount and the nano-sized crystallites. The catalytic performance of methanol-to-aromatics (MTA) indicates that the Z5-0.12 catalyst exhibited higher catalytic activity and selectivity of BTX as the reaction was prolonged, which could result from the synergistic effect among the proper strong acid amount, the smaller zeolite grain size, and the abundant hierarchical structure.

Background: Nitrate reductase (NR) is an important enzyme for nitrate assimilation in plants, and its activity is regulated by post-translational phosphorylation. The change of nitrogen uptake affects the response of rice to low temperature and its growth. To investigate the effect of NIA1 protein dephosphorylation on the growth of rice and its adaptability to low temperature, we analyzed phenotype, chlorophyll content, nitrogen utilization, and antioxidant capacity at low temperature in lines with a mutated NIA1 phosphorylation site (S532D and S532A), an OsNia1 over-expression line (OE), and wild-type Kitaake rice (WT). Results: Plant height, dry matter weight, and chlorophyll content of S532D and S532A were lower than those of WT and OE under normal growth conditions but were higher than those of WT and OE at low temperature. Compared with WT and OE, the nitrite, H2O2, and MDA contents of S532D and S532A leaves were higher under normal growth conditions. The difference in leaf nitrite content between transgenic lines and WT was narrower at low temperature, especially in S532D and S532A, while H2O2 and MDA contents of S532D and S532A leaves were lower than those in WT and OE leaves. The NH4+-N and amino acid contents of S532D and S532A leaves were higher than those of WT and OE leaves under normal or low temperature. qRT-PCR results revealed that transcription levels of OsNrt2.4, OsNia2, and OsNADH-GOGAT were positively correlated with those of OsNia1, and the transcription levels of OsNrt2.4, OsNia2, and OsNADH-GOGAT were significantly higher in transgenic lines than in WT under both normal and low temperature. Conclusions: Phosphorylation of NR is a steady-state regulatory mechanism of nitrogen metabolism, and dephosphorylation of NIA1 protein improved NR activity and nitrogen utilization efficiency in rice. Excessive accumulation of nitrite under normal growth conditions inhibits the growth of rice; however, accumulation of nitrite is reduced at low temperature, enhancing the cold tolerance of rice. These results provide a new insight for improving cold tolerance of rice.

It is important to ensure food safety to study the technology and mechanism of pesticide residues degradation in crops. Though γ-aminobutyric acid (GABA) has been widely reported to involve in plant stress resistance, whether exogenous application or endogenous regulation of GABA by gene-editing technology can promote the pesticide detoxification is not clear in plants. Using tomato and chlorothalonil (CHT) as research models, we discovered that exogenous application of GABA or endogenous elevation of GABA by knockout of pyruvate-dependent GABA transaminase promoted both CHT metabolism and plant stress tolerance to CHT. This is closely related to the active adaptation of GABA to CHT stress by regulating the plant GABA shunt pathway and polyamine pathway. The transcriptome data revealed 17 target genes that may be closely related to the involvement of GABA in CHT metabolism, including 4 peroxidases, 5 glycosyltransferases, 4 glutathione S-transferases, and 4 ABC transporters. In addition, the glutathione detoxification pathway and antioxidative enzyme also actively participated in the GABA-induced CHT detoxification process, which played an important role in relieving CHT stress. As a result, GABA significantly increased the photosynthetic capacity of tomato leaves under CHT stress. While studying photosynthesis, we unexpectedly found that GABA promotes stomatal closure in terms of decreased stomatal conductance and stomatal diameter. This result implies that GABA can reduce CHT absorption by regulating stomatal movement in leaves. Together, we provided a novel viewpoint that foliar application of GABA or metabolic engineering of GABA is an effective approach to reduce the risk of pesticide contamination in crop production.

Multiple morphological abnormalities of the flagella (MMAF) can lead to male infertility due to impaired sperm motility and morphology. Calmodulin- and spoke-associated complex (CSC) are known for their roles in radial spoke (RS) assembly and ciliary motility in Chlamydomonas, while the role of cilia- and flagella-associated protein 61 (CFAP61), a mammalian ortholog of the CSC subunits, in humans is yet unknown. Here, we recruited three unrelated Pakistani families comprising of 11 infertile male patients diagnosed with MMAF. CFAP61 variants, c.451_452del (p.I151Nfs*4) in family 1 and c.847C > T (p.R283*) in family 2 and 3, were identified recessively co-segregating with the MMAF phenotype. Transmission electron microscopy analyses revealed severe disorganized axonemal ultrastructures, and missings of central pair, RSs, and inner dynein arms were also observed and confirmed by immunofluorescence staining in spermatozoa from patients. CFAP61 and CFAP251 signals were absent from sperm tails of the patients, which suggested the loss of functional CSC in sperm flagella. Altogether, our findings report that homozygous variants in CFAP61 are associated with MMAF and male infertility, demonstrating the essential role of this gene in normal sperm flagellum structure in humans.

It is estimated that approximately 25% of nonobstructive azoospermia (NOA) cases are caused by single genetic anomalies, including chromosomal aberrations and gene mutations. The identification of these mutations in NOA patients has always been a research hot spot in the area of human infertility. However, compared with more than 600 genes reported to be essential for fertility in mice, mutations in approximately 75 genes have been confirmed to be pathogenic in patients with male infertility, in which only 14 were identified from NOA patients. The small proportion suggested that there is much room to improve the methodology of mutation screening and functional verification. Fortunately, recent advances in whole exome sequencing and Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)–Cas9 have greatly promoted research on the etiology of human infertility and made improvements possible. In this review, we have summarized the pathogenic mutations found in NOA patients and the efforts we have made to improve the efficiency of mutation screening from NOA patients and functional verification with the application of new technologies.

Description, RAD51AP2 is required specifically for efficient meiotic recombination to form crossover between X and Y chromosomes., Faithful segregation of X and Y chromosomes requires meiotic recombination to form a crossover between them in the pseudoautosomal region (PAR). Unlike autosomes that have approximately 10-fold more double-strand breaks (DSBs) than crossovers, one crossover must be formed from the one or two DSBs in PARs, implying the existence of a sex chromosome–specific recombination mechanism. Here, we found that RAD51AP2, a meiosis-specific partner of RAD51, is specifically required for the crossover formation on the XY chromosomes, but not autosomes. The decreased crossover formation between X and Y chromosomes in Rad51ap2 mutant mice results from compromised DSB repair in PARs due to destabilization of recombination intermediates rather than defects in DSB generation or synapsis. Our findings provide direct experimental evidence that XY recombination may use a PAR-specific DSB repair mechanism mediated by factors that are not essential for recombination on autosomes.

Carotenoids are the main pigments responsible for the coloration and account for the major antioxidant activity of tomato (Solanum lycopersicum L.) fruit. Significant increments in total carotenoids and lycopene levels were observed in tomato fruit illuminated by red light relative to white light in previous studies, but the mechanism of carotenoid biosynthesis regulated by red light is still unclear. In the present study, the influence of red light on carotenoid biosynthesis in postharvest tomato fruit was conducted using targeted metabolomics and transcriptomic methods. A total of 25 differentially accumulated carotenoids and 1939 differentially expressed genes were isolated and identified. The results illustrated that the contents of phytoene and lycopene were considerably higher in fruit treated with red light than those with white light at 12 h. These differentially expressed genes are mainly enriched in plant hormone signal transduction, photosynthesis, secondary metabolite biosynthesis, and plant circadian rhythm. Moreover, from the results of coexpression network analysis, 15 transcription factors from red light-treated fruit were screened; among these, transcription factors of SlERF4, SlbHLH93 and SlIAA29, which are involved in signal transduction of light and hormones, respectively, may also play important roles in carotenoid biosynthesis regulated by red light in tomato fruit. It is concluded that red light enhanced carotenoid biosynthesis in postharvest tomato fruit and the mechanisms of enhanced carotenoid biosynthesis were not only associated with the direct regulation by red light signalling, but also with the indirect regulation by hormonal signalling.

In this study, an integrated transcriptome and metabolome analysis was used to explore the molecular foundation of fruit quality in two parent lines of Citrullus lanatus with distinct flesh characteristics, including ‘14-1’ (sweet, red, and soft) and ‘W600’ (bitter, light yellow, and firm), as well as the corresponding F1 population (bitter, light yellow, and firm). Numerous differentially expressed genes (DEGs) were identified in the fruit samples: 3766 DEGs for ‘14-1’ vs. ‘W600’, 2767 for ‘14-1’ vs. F1, and 1178 for F1 vs. ‘W600’ at the transition stage; and 4221 for ‘14-1’ vs. ‘W600’, 2447 for ‘14-1’ vs. F1, and 446 for F1 vs. ‘W600’ at the maturity stage. Weighted gene co-expression network analysis (WGCNA) revealed that a gene module including 1111 DEGs was closely associated with flesh taste and color, and another gene module including 1575 DEGs contributed significantly to flesh texture. The metabolomic results showed that there were 447 differential metabolites (DMs) for ‘14-1’ vs. ‘W600’ fruits, 394 for ‘14-1’ vs. F1, and 298 for F1 vs. ‘W600’ at the maturity stage. Combining WGNCA and metabolomic results, several DEGs and DMs were further identified as hub players in fruit quality formation: 6 DEGs with 4 DMs for flesh sweetness; 6 DEGs with 13 DMs for bitterness; 9 DEGs with 10 DMs for flesh color; and 9 DEGs with 4 DMs for flesh texture. Altogether, these observations not only expand our knowledge of the molecular basis of fruit quality in watermelon, but also provide potential targets for future watermelon improvement.

The low absolute positioning accuracy of industrial robots is one of the bottlenecks preventing industrial robots from precision applications. Kinematic calibration is the main way to improve the absolute positioning accuracy of industrial robots, which greatly relies on three-dimensional (3D) measurement instruments, including laser trackers and pull rope mechanisms. These instruments are costly, and their required intervisibility space is large. In this paper, a precision 3D measurement instrument integrating multiple laser range sensors is designed, which fuses the information of multiple redundant laser range sensors to obtain the coordinates of a 3D position. An identification model of laser beam position and orientation parameters based on redundant distance information and standard spherical constraint is then developed to reduce the requirement for the assembly accuracy of laser range sensors. A hybrid identification algorithm of PSO-LM (particle swarm optimization Levenberg Marquardt) is designed to solve the high-order nonlinear problem of the identification model, where PSO is used for initial value identification, and LM is used for final value identification. Experiments of identification of position and orientation, verifications of the measuring accuracy, and the calibration of industrial robots are conducted, which show the effectiveness of the proposed 3D measurement instrument and identification methods. Moreover, the proposed instrument is small in size and can be used in narrow industrial sites.

Melatonin (MT) is a phytohormone-like substance and is profoundly involved in modulating nearly all aspects of plant development and acclimation to environmental stressors. However, there remain no studies about the effects of MT on tomato seed germination under salt stress. Here we reported that the overexpression of caffeic acid O-methyltransferase 1 (SlCOMT1) significantly increased both MT content and salt tolerance in the germinated seeds of a transgenic tomato relative to wild type (WT) samples. Physiological investigation showed higher amylase activity in the stressed overexpression seeds than WT, leading to the promoted starch decomposition and enhanced soluble sugar content. The stimulated production of osmolytes and enhanced activities of SOD, POD, and CAT, together with the significant reduction in H2O2 and O2·− accumulation, were revealed in the stressed overexpression seeds relative to WT, largely accounting for their lower membrane lipid peroxidation. qPCR assay showed that, upon salt stress, the transcript abundance of hub genes related to germination (SlCYP707A1, SlABA1, SlGA3ox2 and SlGA2ox4) and stress tolerance (SlCDPK1, SlWRKY33 and SlMAPK1) were distinctly altered in the overexpression samples when compared to WT, providing a molecular basis for MT-mediated improvement of seed salt tolerance. Altogether, our observations shed new insights into biological functions of SlCOMT1 and could expand its utilization in genetic improvement of tomato salt tolerance in future.

Preantral follicles are often used as models for cryopreservation and in vitro culture due to their easy availability. As a promising approach for mammalian fertility preservation, vitrification of preantral follicles requires high concentrations of highly toxic penetrating cryoprotective agents (up to 6 M). Here, we accomplish low-concentration-penetrating cryoprotective agent (1.5 M) vitrification of mouse preantral follicles encapsulated in hydrogel by nanowarming. We find that compared with conventional water bath warming, the viability of preantral follicles is increased by 33%. Moreover, the cavity formation rate of preantral follicles after in vitro culture is comparable to the control group without vitrification. Furthermore, the percentage of MII oocytes developed from the vitrified follicles, and the birth rate of offspring following in vitro fertilization and embryo transfer are also similar to the control group. Our results provide a step towards nontoxic vitrification by utilizing the synergistic cryoprotection effect of microencapsulation and nanowarming.

Multiple morphological abnormalities of the flagella (MMAF) can lead to male infertility due to impaired sperm motility and morphology. Calmodulin- and spoke-associated complex (CSC) are known for their roles in radial spoke (RS) assembly and ciliary motility in Chlamydomonas, while the role of cilia- and flagella-associated protein 61 (CFAP61), a mammalian ortholog of the CSC subunits, in humans is yet unknown. Here, we recruited three unrelated Pakistani families comprising of 11 infertile male patients diagnosed with MMAF.