Decreased LONP1 expression contributes to DNA damage and meiotic defects in oocytes.

Meiotic defects in oocytes are the primary reason for decreased female fertility with advanced maternal age. In this study, we revealed that decreased expression of ATP-dependent Lon peptidase 1 (LONP1) in aged oocytes and oocyte-specific depletion of LONP1 disrupt oocyte meiotic progression accompanying with mitochondrial dysfunction. In addition, LONP1 downregulation increased oocyte DNA damage. Moreover, we demonstrated that splicing factor proline and glutamine rich directly interacts with LONP1 and mediate the effect of LONP1 depletion on meiotic progression in oocytes. In summary, our data suggest that decreased expression of LONP1 is involved in advanced maternal age-related meiosis defects and that LONP1 represents a new therapeutic target to improve aged oocyte quality.

Downregulated INHBB in endometrial tissue of recurrent implantation failure patients impeded decidualization through the ADCY1/cAMP signalling pathway.

PURPOSE: This study aims to identify the mechanism of Inhibin Subunit Beta B (INHBB), a member of the transforming growth factor-ß (TGF-ß) family involved in the regulation of human endometrial stromal cells (HESCs) decidualization in recurrent implantation failure (RIF). METHODS: RNA-seq was conducted to identify the differentially expressed genes in the endometria from control and RIF patients. RT-qPCR, WB, and immunohistochemistry were performed to analyse the expression levels of INHBB in endometrium and decidualised HESCs. RT-qPCR and immunofluorescence were used to detect changes in the decidual marker genes and cytoskeleton after knockdown INHBB. Then, RNA-seq was used to dig out the mechanism of INHBB regulating decidualization. The cAMP analogue (forskolin) and si-INHBB were used to investigate the involvement of INHBB in the cAMP signalling pathway. The correlation of INHBB and ADCY expression was analysed by Pearson's correlation analysis. RESULTS: Our results showed significantly reduced expression of INHBB in endometrial stromal cells of women with RIF. In addition, INHBB was increased in the endometrium of the secretory phase and significantly induced in in-vitro decidualization of HESCs. Notably, with RNA-seq and siRNA-mediated knockdown approaches, we demonstrated that the INHBB-ADCY1-mediated cAMP signalling pathway regulates the reduction of decidualization. We found a positive association between the expression of INHBB and ADCY1 in endometria with RIF (R2 = 0.3785, P = 0.0005). CONCLUSIONS: The decline of INHBB in HESCs suppressed ADCY1-induced cAMP production and cAMP-mediated signalling, which attenuated decidualization in RIF patients, indicating that INHBB is an essential component in the decidualization process.

First report of peanut root rot caused by Fusarium acuminatum in Shandong Province, China.

Peanut (Arachis hypogaea L.) is one of the most economically important crops as a major source of edible oil and protein. In July 2021, a root rot disease was observed on peanut in Laiwu (36º22' N, 117º67' E), Shandong Province, China. Disease incidence was approximately 35%. Disease symptoms included root rot, vessels with a brown to dark brown discoloration, plus progressive yellowing and wilting of leaves from the base leading to whole plant death. To determine the causal agent, symptomatic roots with typical lesions were cut into small pieces, surface sterilized in 75% ethanol for 30 s, and 2% NaClO for 5 min, rinsed three times in sterile water and placed on potato dextrose agar (PDA) at 25℃ (Leslie and Summerell 2006). After 3 days of incubation, whitish-pink to red colonies growing from the roots were observed. Eight single-spore isolates had identical morphological traits that were similar to those of Fusarium spp. A representative isolate (LW-5) was used for morphological characterization, molecular analysis, and pathogenicity test. On PDA, the isolate formed dense aerial mycelia, which were initially white, then became deep pink with age and formed red pigments in the medium. On carnation leaf agar (CLA), macroconidia with 3 to 5 septa were abundant, relatively slender, curved to lunate, that measured 23.7 to 52.2 × 3.6 to 5.4 µm (n=50). Microconidia were oval, 0 to 1 septa. Chlamydospores were globose with a smooth outer wall in chains or single. Following DNA extraction of isolate LW-5, primers EF1-728F/EF1-986R (Carbone et al., 1999), RPB1U/RPB1R, and RPB2U/RPB2R (Ponts et al., 2020) were used to amplify the partial translation elongation factor 1 alpha (TEF1-α), RNA polymerase II largest subunit (RPB1), and RNA polymerase II second largest subunit (RPB2) regions for DNA sequencing, respectively. BLASTn analysis of TEF1-α (GenBank accession No. OP838084), RPB1 (OP838085), and RPB2 (OP838086) sequences, revealed 99.66, 99.87, and 99.09% identity with those of F. acuminatum (OL772800, OL772952 and OL773104), respectively. Isolate LW-5 was identified as F. acuminatum based on morphology and molecular analysis. Twenty Huayu36 peanut seeds were each planted in a 500-ml sterile pot containing 300 g of autoclaved potting medium (nutritive soil: vermiculite=2:1 in volume). Two weeks after seedling emergence, 1 cm depth of the potting medium was dug around the plants to expose the taproot. Two 5-mm wounds per taproot were scratched with a sterile syringe needle. Potting medium in each pot of 10 inoculated plants was mixed with 5 ml of conidial suspension (106 conidia per ml). The other 10 plants were used as non-inoculated controls and treated with sterile water in the same manner. The seedlings were placed in a plant growth chamber maintained at 25°C, RH >70%, 16-h light per day, and irrigated with sterile water. After 4 weeks, inoculated plants exhibited yellowing and wilting symptoms that were similar to those observed in the field, while non-inoculated control plants had no symptoms. F. acuminatum was re-isolated from diseased roots and confirmed using morphological features and DNA sequence analysis of TEF1-α, RPB1 and RPB2. F. acuminatum was reported to cause root rot on Ophiopogon japonicus (Linn. f.) (Tang et al., 2020), Polygonatum odoratum (Li et al., 2021), and Schisandra chinensis (Shen et al., 2022) in China. To our knowledge, this is the first report of root rot on peanut caused by F. acuminatum in Shandong Province, China. Our report will provide crucial information for studying the epidemiology and management of this disease.

First Report of Colletotrichum gloeosporioides Causing Anthracnose on Peanut in Chongqing, China.

In July 2022, dieback and discoloration were detected on infected stems of peanut in Qijiang District of Chongqing (106.56°E,29.41°N), China, with an incidence up to 5%. These peanut stems had disease symptoms typical of anthracnose with irregular gray-brown spots with dark brown edges, sunken, and necrotic. High temperature and high humidity were favorable for the growth of the pathogen. To isolate the pathogen, we collected 10 typical infected peanuts and cut one piece from each of symptomatic stems, surface sterilized with 0.5% NaClO for 1 min, and 75% ethanol for 30 s, then rinsed three times with sterile distilled water and dried on sterilized filter paper. These pieces were incubated on potato dextrose agar (PDA) at 25°C in the dark. Pure cultures were obtained from hyphal tips of each colony. It was found that isolates with the same colony morphology were isolated from each infected stem. A representative isolate (L7) was used for morphological characterization, molecular analysis, phylogenetic analysis, and pathogenicity tests. The colonies appeared white to gray, with white margins and aerial hyphae, and the reverse of the colonies was gray to brown. Conidia were cylindrical, aseptate, with obtuse to slightly rounded ends, 13.4 to 18.8 × 4.2 to 5.8 µm (n=50). Morphological characteristics were generally consistent with those of Colletotrichum gloeosporioides species complex (Cannon et al., 2012). For molecular identification, genomic DNA was extracted using a CTAB method and partial sequences of ß-tubulin (TUB2), actin (ACT) genes, chitin synthase (CHS) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) genes were amplified and sequenced using primers T1/T2, ACT-512F/ACT-783R, CHS-79F/CHS-345R, and GDF1/GDR1, respectively (Damm et al., 2012; Dowling et al., 2020). Using the BLAST, TUB2, ACT, CHS and GAPDH gene sequences (GenBank accession No. OR714793, OP168707, OP168708 and OR714794, respectively) were100% (429 bp out of 429 bp), 99.22% (256 bp out of 258 bp), 99.64% (276 bp out of 277 bp) and 100% (253 bp out of 253 bp) identical to C. gloeosporioides CBS:112999 (JQ005587, JQ005500, JQ005326, and JQ005239), respectively. Using Neighbor-Joining algorithm, phylogenetic analysis was conducted based on the concatenated sequences of published TUB2, ACT, CHS and GAPDH genes. The identified isolate (L7) was closely related to C. gloeosporioides. To evaluate pathogenicity, the stems of ten peanut (Zhonghua12) seedlings (2 weeks) were wounded with a sterile toothpick and mycelial plugs (5 mm in diameter) or 20 µl of conidial suspension (105/ml) were inoculated. Non-colonized agar plugs or 20 µl of sterile distilled water were treated as control. After inoculation, the peanuts were kept in a moist chamber at 28°C with 80% humidity in the dark for 24 h, and subsequently transferred to the moist chamber with 12 h light and darkness cycle for 6 days, similar symptoms were observed on all inoculated peanuts. Controls remained asymptomatic. C. gloeosporioides was reisolated from the diseased stems and confirmed using morphological features and sequence analysis of TUB2, ACT, CHS and GAPDH. Anthracnose caused by C. truncatum and C. fructicola has been reported on peanut leaves in China (Gong et al., 2023; Yu et al., 2019). To our knowledge, this is the first report of anthracnose on peanut stem caused by C. gloeosporioides in Chongqing. Our report will provide crucial information for studying on epidemiology and management of this disease.

First Report of Root Rot Caused by Macrophomina phaseolina in Peanut Shandong Province, China.

Peanut (Arachis hypogaea L.) is a widely grown oilseed crop of great agricultural importance worldwide. In July 2022, disease symptoms were observed on peanut roots in Laixi (36º85' N, 120º54' E), Shandong Province, China. About 25% of the plants showed various symptoms, including stem and root rot and blackening, microsclerotia on the stem, yellowing and wilting of leaves, and even death. Twenty diseased plants were collected to confirm the pathogen. Symptomatic roots were cut into small pieces, disinfested with 75% ethanol for 1 min and 0.5% NaClO for 2 min, rinsed three times with sterile water, dried on sterile filter paper, and then spread on potato dextrose agar (PDA) supplemented with 100 µg/mL chloramphenicol and incubated at 25°C in the dark. At the beginning of growth, the fungus formed sparse, white mycelia, which white, then darkened with age and microsclerotia were formed in the medium after 5 days. The mycelium aggregated into black, round to oblong or irregularly shaped microsclerotia 84 to 163 µm long and 54 to 125 µm wide (n=40). These morphological characteristics were consistent with the description of Macrophomina phaseolina (Holliday and Punithalingam, 1970). Molecular identification was performed by sequencing the internal transcribed spacer (ITS) region with ITS1 and ITS4 and translation elongation factor 1-alpha (TEF) with EF1-728F/EF1-986R (Glass and Donaldson 1995) of a representative isolate SXY183. ITS (OR056369) and TEF (OR098356) of SXY183 showed 100% and 97.74% similarity with M. phaseolina (KF951622, KF951997), respectively. Phylogenetic analysis was performed using Neighbor-Joining (NJ) analysis based on the gene sequences of ITS and TEF. The fungus was identified as M. phaseolina based on molecular analysis and morphological characteristics. The pathogenicity of a representative isolate (SXY183) was tested on peanuts under greenhouse conditions. Two-week-old peanut (Huayu No. 9115) seedlings were inoculated with a mycelial plug (8 mm diameter) at the root base of each plant and cultured in a greenhouse (30°C during the day and 25°C at night, a 12-h photoperiod, and 80% RH). Ten plants were inoculated with a plug of non-colonized PDA as a control. Brown lesions were observed on the stem and root of all inoculated seedlings 7 days after inoculation, but not on the control plants. The experiment was repeated three times. M. phaseolina was re-isolated from the symptomatic root and confirmed based on morphological characteristics and DNA sequence analysis of ITS and TEF. M. phaseolina is a soil-borne fungus that is distributed worldwide and has a broad host range. Disease agent has previously been reported on several host plants such as adzuki bean, faba bean, watermelon, Plukenetia volubilis, Atractylodes lancea and Curcuma longa in China (Cai et al., 2020; Sun et al. 2016; Sun et al., 2019; Sun et al., 2020; Wang et al., 2020; Wu et al., 2022). However, this is the first report in which M. phaseolina was found to cause peanut root rot in Shandong Province, China. Our report will provide important information for studying the epidemiology and management of this disease.

Association between gestational trophoblastic disease (GTD) history and clinical outcomes in in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) cycles.

BACKGROUND: Gestational trophoblastic disease (GTD) usually affects young women of childbearing age. After treatment for GTD, 86% of women wish to achieve pregnancy. On account of the impacts of GTD and treatments as well as patient anxiety, large numbers of couples turn to assisted reproductive technology (ART), especially in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI). But few studies have investigated whether a history of GTD affects the outcomes of IVF/ICSI in secondary infertile patients and how it occurs. We investigate whether a history of GTD affects the IVF/ICSI outcomes and the live birth rates in women with secondary infertility. METHODS: This retrospective cohort study enrolled 176 women with secondary infertility who underwent IVF/ICSI treatment at the reproductive medical center of Nanjing Drum Tower Hospital from January 1, 2016, to December 31, 2020. Participants were divided into the GTD group (44 women with GTD history) and control group (132 women without GTD history matched from 8318 secondary infertile women). The control group and the study group were matched at a ratio of 3:1 according to patient age, infertility duration, number of cycles and body mass index (BMI). We assessed retrieved oocytes and high-grade embryos, biochemical pregnancy, miscarriage, ectopic pregnancy, gestational age at delivery, delivery mode and live birth rates. RESULT(S): We found a significantly reduced live-birth rate (34.1% vs 66.7%) associated with IVF/ICSI cycles in patients with a GTD history compared to those without a GTD history. The biochemical pregnancy and miscarriage rates of the GTD group were slightly higher than those of the control group. In addition, there was a difference in gestational age at delivery between the GTD and control groups (p < 0.001) but no differences in the mode of delivery (p = 0.267). Furthermore, the number of abandoned embryos in the GTD group was greater than that in the control group (p = 0.018), and the number of good-quality embryos was less than that in the control group (p = 0.019). The endometrial thickness was thinner (p < 0.001) in the GTD group. Immunohistochemistry (IHC) showed abnormal endometrial receptivity in the GTD group. CONCLUSION(S): The GTD history of patients undergoing IVF/ICSI cycles had an impact on the live-birth rate and gestational age at delivery, which might result from the thinner endometrium and abnormal endometrial receptivity before embryo transfer.

First Report of Nigrospora aurantiaca Causing Leaf Blight Disease of Peanut in China.

In June 2021, a previously unreported leaf blight disease of peanut (Arachis hypogaea) was observed on field-grown peanut (Jinhua19) in Laixi city, Shandong province of China. Approximately 5% of plants showed disease symptoms in the fields we investigated. The symptoms first appeared as yellow round or irregular spots on leaves, and then the spots became brown. As the disease progressed, spots became larger and even converge, which later produced leaf chlorosis and abscission. Symptomatic leaves were cut into small pieces, surface disinfested with 70% ethanol for 30s, 1% NaClO for 60s, rinsed three times in sterile water, dried on sterile filter papers, placed on potato dextrose agar (PDA) media, and incubated at 25°C in darkness. Fungal cultures were initially white, with red pigment, then turned gray, and eventually turned black, and aerial hyphae were dense. Conidia were spherical or slightly ellipsoidal, black, smooth, and 8.6 to 11.5 × 8.7 to 14.5µm (n=50). Morphological characteristics of the isolates matched the description of Nigrospora aurantiaca (Wang et al. 2017). Molecular identification was performed by sequencing beta tubulin gene (TUB) with Bt2a/Bt2b and translation elongation factor 1-alpha (TEF) with EF1-728F/EF1-986R (Wang et al. 2021) of a representative isolate ZHX11. TUB (OK489789) and TEF (OK489790) of ZHX11 obtained 100% (401/401 nucleotides) and 99.64% (279/279 nucleotides) similar to those of N. aurantiaca (MN329935, MN264010), respectively. Alignment was conducted separately for each gene set using Clustal W algorithm implemented in MEGA 7.0 (Kumar et al. 2016), and multi-gene (TUB and TEF) phylogenetic analyses using Neighbor-Joining (NJ) method showed that the isolate was N. aurantiaca. To complete Koch's postulates, nine 2-week-old peanut (Zhonghua 12) seedlings were sprayed with conidia suspensions (106 conidia mL-1 in 0.05% Tween 20 buffer). The same number of seedlings were only treated with 0.05% Tween buffer as controls. The experiment was repeated three times. Plants were incubated in a growth chamber (30°C in the day and 25°C at night, a 12-h photoperiod and 80% RH). Ten days after inoculation, typical symptoms were observed on inoculated leaves but not on the controls. N. aurantiaca was reisolated from the diseased leaves but not from the controls. N. sphaerica was observed on peanut in China (Liu et al. 2020). To our knowledge, this is the first report of N. aurantiaca causing leaf blight on peanut in shandong province, China. These findings will help to develop better preventive measures in accordance with the emergence of the new disease.

Multi-Gas Detection System Based on Non-Dispersive Infrared (NDIR) Spectral Technology.

Automobile exhaust gases, such as carbon dioxide (CO2), carbon monoxide (CO), and propane (C3H8), cause the greenhouse effect, photochemical smog, and haze, threatening the urban atmosphere and human health. In this study, a non-dispersive infrared (NDIR) multi-gas detection system consisting of a single broadband light source, gas cell, and four-channel pyroelectric detector was developed. The system can be used to economically detect gas concentration in the range of 0-5000 ppm for C3H8, 0-14% for CO, and 0-20% for CO2. According to the experimental data, the concentration inversion model was established using the least squares between the voltage ratio and the concentration. Additionally, the interference coefficient between different gases was tested. Therefore, the interference models between the three gases were established by the least square method. The concentration inversion model was experimentally verified, and it was observed that the full-scale error of the sensor changed less than 3.5%, the detection repeatability error was lower than 4.5%, and the detection stability was less than 2.7%. Therefore, the detection system is economical and energy efficient and it is a promising method for the analysis of automobile exhaust gases.

First comprehensive proteomics analysis of lysine crotonylation in leaves of peanut (Arachis hypogaea L.).

Lysine crotonylation is an important post-translational modification process. Most research in this area has been carried out on mammals and yeast, but there has been little research on it in plants. In the current study, large-scale lysine crotonylome analysis was performed by a combination of affinity enrichment and high-resolution LC-MS/MS analysis. Altogether, 6051 lysine crotonylation sites were identified in 2508 protein groups. Bioinformatics analysis showed that lysine-crotonylated proteins were involved in many biological processes, such as carbon fixation in photosynthetic organisms, biosynthesis of amino acids, ribosomes structure and function. In particular, subcellular localization analysis showed that 43% of the crotonylated proteins were located in the chloroplast. Twenty-nine crotonylation proteins were associated with photosynthesis and functional enrichment that these proteins were associated with the reaction center, photosynthetic electron transport, and ATP synthesis. Based on these results, further studies to expand on the lysine crotonylome analysis were suggested.

Microbial metabolism and adaptations in Atribacteria-dominated methane hydrate sediments.

Gas hydrates harbour gigatons of natural gas, yet their microbiomes remain understudied. We bioprospected 16S rRNA amplicons, metagenomes, and metaproteomes from methane hydrate-bearing sediments under Hydrate Ridge (offshore Oregon, USA, ODP Site 1244, 2-69 mbsf) for novel microbial metabolic and biosynthetic potential. Atribacteria sequences generally increased in relative sequence abundance with increasing sediment depth. Most Atribacteria ASVs belonged to JS-1-Genus 1 and clustered with other sequences from gas hydrate-bearing sediments. We recovered 21 metagenome-assembled genomic bins spanning three geochemical zones in the sediment core: the sulfate-methane transition zone, the metal (iron/manganese) reduction zone, and the gas hydrate stability zone. We found evidence for bacterial fermentation as a source of acetate for aceticlastic methanogenesis and as a driver of iron reduction in the metal reduction zone. In multiple zones, we identified a Ni-Fe hydrogenase-Na+ /H+ antiporter supercomplex (Hun) in Atribacteria and Firmicutes bins and in other deep subsurface bacteria and cultured hyperthermophiles from the Thermotogae phylum. Atribacteria expressed tripartite ATP-independent transporters downstream from a novel regulator (AtiR). Atribacteria also possessed adaptations to survive extreme conditions (e.g. high salt brines, high pressure and cold temperatures) including the ability to synthesize the osmolyte di-myo-inositol-phosphate as well as expression of K+ -stimulated pyrophosphatase and capsule proteins.

First Report of Fusarium ipomoeae Causing Peanut leaf Spot in China.

Peanut (Arachis hypogaea L.) is one of the most economically important crops as an important source of edible oil and protein. In August 2020, circular to oval-shaped brown leaf spots (2-6 mm in diameter) with well-defined borders surrounded by a yellow margin were observed on peanut plant leaves in Laixi City, Shandong Province, China. Symptomatic plants randomly distributed in the field, the incidence was approximately 5%. Leave samples were collected consisted of diseased tissue and the adjacent healthy tissue. The samples were dipped in a 70% (v/v) ethanol solution for 30 s and then soaked in a 0.1% (w/v) mercuric chloride solution for 60 s. The surface-sterilized tissues were then rinsed three times with sterile distilled water, dried and placed on Czapek Dox agar supplemented with 100 µg/ml of chloramphenicol. The cultures were incubated in darkness at 25 °C for 3-5 days. Fungal colonies were initially white and radial, turning to orange-brown in color, with abundant aerial mycelia. Macroconidia were abundant, 4 to 7 septate, with a dorsiventral curvature, and were 3.3-4.5 × 18.5-38.1 µm (n=100) in size; microconidia were absent; chlamydospores were produced in chains or clumps, ellipsoidal to subglobose, and thick walled. The morphological characteristics of the conidia were consistent with those of Fusarium spp. To identify the fungus, an EasyPure Genomic DNA Kit (TransGEN, Beijing, China) was used to extract the total genomic DNA from mycelia. The internal transcribed spacer region (ITS rDNA) and the translation elongation factor 1-α gene (TEF1) were amplified with primers ITS1/ITS4 (White et al. 1990) and EF1/EF2 (O'Donnell et al. 1998), respectively. Based on BLAST analysis, sequences of ITS (MT928727) and TEF1 (MT952337) showed 99.64% and 100% similarity to the ITS (MT939248.1), TEF1 (GQ505636.1) of F. ipomoeae isolates. Sequence analysis confirmed that the fungus isolated from the infected peanut was F. ipomoeae (Xia et al. 2019). The pathogenicity of the fungus was tested in the greenhouse. Twenty two-week-old peanut seedlings (cv. Huayu20) grown in 20-cm pots (containing autoclaved soil) were sprayed with a conidial suspension (105 ml-1) from a 15-day-old culture. Control plants were sprayed with distilled water. The experiment was conducted as a randomized complete block design, and placed at 25 °C under a 12-h photoperiod with 90% humidity. Symptoms similar to those in the field were observed on leaves treated with the conidial suspension ten days after inoculation, but not on control plants. F. ipomoeae was re-isolated from symptomatic leaves but not from the control plants. Reisolation of F. ipomoeae from inoculated plants fulfilled Koch's postulates. To our knowledge, this is the first report of F. ipomoeae causing peanut leaf spot in China. Our report indicates the potential spread of this pathogen in China and a systematic survey is required to develop effective disease management strategies.

Draft Genome Sequence of Phoma arachidicola Wb2 Causing Peanut Web Blotch in China.

Peanut web blotch, a peanut disease with both web and blotch symptom leaflets, is an emerging threat for peanut cultivation worldwide and one of the most important fungal diseases in China. However, the limited pieces of information in genomic resources and pathogenesis are the major constraints to integrated disease management. The genome contains a large number of pathogenicity-related genes, but the genomic information of the pathogen is still blank. Considering this fact, current study presented the draft genome sequence of a Phoma arachidicola isolate named Wb2. Strain Wb2 was isolated from peanut leaves with typical web blotch symptoms, and identified as Phoma arachidicola based on morphological characteristics and phylogenic analysis using ITS sequence. The draft genome of Wb2 is about 34.11 Mb and contains 37330 open reading frames (ORFs), with G + C content 49.23%. The strain Wb2 has an abundance of secreted oxidases, peroxidases, and carbohydrate-active enzymes for degrading cell wall polysaccharides and penetrating into the host tissue. The genome information of Wb2 will help to better understand the mechanisms of interaction between P. arachidicola and peanuts. Furthermore, the genome-based plant-pathogen interaction analysis will provide clues for disease control, which is essential to ensure peanut production and food security.

Lactate enhanced the effect of parathyroid hormone on osteoblast differentiation via GPR81-PKC-Akt signaling.

Osteoblast uses aerobic glycolysis to meet the metabolic needs in differentiation process. Lactate, the end product of glycolysis, presents in the environment with elevated PTH and osteoblast differentiation. Although previous findings showed that lactate promoted osteoblast differentiation, whether lactate affects PTH-mediated osteoblast differentiation is unclear. To investigate this, pre-osteoblast cell line MC3T3-E1 was treated PTH with or without physiological dose of lactate. Lactate increases ALP positive cell formation, increases ALP activity and expression of differentiation related markers, enriches the CREB transcriptional factor target genes in PTH treated cells. Using inhibitors for MCT-1 reveales that lactate effects are MCT-1 independent. Lactate selectively increases Akt and p38 activation but not Erk1/2 and ß-Catenin activation. The inhibitors for Akt and p38 inhibit lactate effects on PTH mediated osteoblast differentiation. Using inhibitors for Gαi signaling of GPR81 further increases Alp mRNA levels in lactate and PTH co-treatment cells. However, with the inhibitors for Gßγ-PLC-PKC signaling, the effect of lactate on PTH mediated osteoblast differentiation is inhibited. Our data demonstrate that lactate activates GPR81-Gßγ-PLC-PKC-Akt signaling to regulate osteoblast differentiation that mediated by PTH treatment.

Biochar Decreases Fertilizer Leaching and Promotes Miscanthus Growth in Saline-Alkaline Soil.

Biochar has been widely reported to improve soil conditions and affect plant growth. However, its effectiveness is limited by soil type and production technology. Considering the application effect of biochar in saline alkali soil, there is currently a lack of in-depth mechanism explanations in the research. Therefore, we designed an experiment to explore the effect of biochar on plant growth in saline alkali soil and conducted soil column experiments in a greenhouse environment using composite inorganic fertilizer (NPK). The results showed that biochar significantly affected the distribution of soil nutrient content at different depths, with a significant increase in fertility levels in the surface and middle layers and a decrease in fertility levels in deep soils. Compared to using fertilizers alone, the combined use of biochar and fertilizers further expands the enrichment effect and significantly reduces the leaching of fertilizers into deeper layers. At the same time, the application of biochar also improved soil properties, including an increase in electrical conductivity and organic matter content, as well as an increase in soil enzyme activity. On the other hand, the application of biochar also increases the activity of antioxidant enzymes and the content of osmoregulation substances in plants, reducing the environmental stress that plants are subjected to. Therefore, our results indicate that biochar can reduce the leaching of fertilizers into deep soil layers, improve soil properties, and promotes the growth of Miscanthus in saline alkali soils.

Isolation and Identification of Bacillus subtilis LY-1 and Its Antifungal and Growth-Promoting Effects.

Peanut root rot, caused by Fusarium spp., is a devastating fungal disease. As part of a program to obtain a biocontrol agent to control peanut root rot in the field, a bacterial strain LY-1 capable of inhibiting the growth of the fungus in vitro was isolated from rhizosphere soil samples collected from wild mint by agar disk dilution and dual-culture assay. Strain LY-1 was identified as Bacillus subtilis based on morphological characteristics, 16S rDNA, and gyrA sequence analyses. The bacterial suspension and cell-free culture filtrate of LY-1 could significantly inhibit the growth of Fusarium oxysporum, Fusarium proliferatum and Fusarium solani, but volatile organic compounds from the cultures had only a weak effect on mycelial growth. The percentage inhibition of 20% concentration of the cell-free culture filtrate of LY-1 on conidium production of each of the three Fusarium species was greater than 72.38%, and the percentage inhibition by the culture filtration on the germination of conidia of the three species was at least 62.37%. The production of extracellular enzyme activity by LY-1 was studied in functional assays, showing protease, cellulase, amylase, chitinase, and ß-1,3-glucanase activity, while LY-1 contained a gene encoding iturin, an antifungal lipopeptide. In addition, under pot culture in a greenhouse, culture filtrate of LY-1 significantly promoted the growth of peanut, increasing the fresh and dry mass of the plant by 30.77% and 27.27%, respectively, in comparison with the no-filtrate control. The culture filtrate of LY-1 increased the resistance of peanut plants to F. oxysporum, with the biocontrol efficiency reaching 44.71%. In conclusion, B. subtilis LY-1, a plant-growth-promoting rhizobacterium, was able to protect peanuts from Fusarium spp. infection.

Effect of Reducing Nitrogen Fertilization and Adding Organic Fertilizer on Net Photosynthetic Rate, Root Nodules and Yield in Peanut.

Long-term excessive application of chemical fertilizers can cause many problems, such as soil degradation and environmental pollution. Therefore, we reduced conventional nitrogen fertilization and added organic fertilizers in some cases to investigate the response of photosynthetic characteristics, root nodules and yield on reduced nitrogen fertilization. Compared to conventional nitrogen fertilization, the 25% and 35% nitrogen reduction treatments reduced the leaf area index, net photosynthetic rate, 100-fruit weight, 100-kernel weight and the yield of peanut, but had no significant effect on the kernel rate. With constant N fertilizer, adding organic fertilization alone increased leaf area index, chlorophyll, net photosynthetic rate and yield of peanut. In compounded treatments of nitrogen and organic fertilizer, the highest yields were achieved in the 25% N reduction with the 3000 kg/hm-2 organic fertilizer treatment (T3) and the 4500 kg/hm-2 organic fertilizer treatment (T4); furthermore, the net photosynthetic rate, leaf area index, yield and fertilizer contribution were significantly higher in these two treatments than in the conventional fertilizer treatments. Nitrogen fertilizer had significant effects on the quantity and fresh weight of root nodules. Concretely, nitrogen reduction increased the quantity and fresh weight of root nodules of peanut in the early stage of fertility but decreased them in the harvest stage. Nitrogen reduction with an additional organic fertilizer in the late stage of fertility increased the quantity and fresh weight of root nodules of peanut. Considering the property of root nodules was significantly positively correlated with net photosynthetic rate and yield, the arguments above may be the mechanism of the highest yields found in T3 and T4. This work can provide empirical and instructional support for a balanced fertilization strategy in peanut agriculture and high-yielding and efficient cultivation of peanut.

Arbuscular Mycorrhizal Fungi Restored the Saline-Alkali Soil and Promoted the Growth of Peanut Roots.

Peanut (Arachis hypogaea L.) is an important oil and cash crop. An efficient utilization of saline-alkali soil resources, the development of peanut planting in saline-alkali soil, and obtaining high and stable yield have become urgent needs to ensure peanut production. Arbuscular mycorrhizal fungi (AMF) have been reported to develop the potential productivity of host plants and improve their salt resistance and tolerance. However, there is still limited research on promoting the growth and morphology of peanut roots. Therefore, in this study, seeds of salt-tolerant peanut variety "HY 25" were coated with commercial AMF inoculant before being planted in saline-alkali and normal soils to investigate the effects of AMF on peanut root growth and rhizosphere soil. The results showed that root morphological characteristics were significantly increased by the use of AMF at the podding stage in saline-alkali soil and from the flowering and pegging stage to the maturity stage in normal soil. Of note, the total root volume of peanut inoculated with AMF significantly increased by 31.57% during the podding stage in saline-alkali soil. Meanwhile, AMF significantly increased the phosphatase and invertase activities in the peanut rhizosphere of saline-alkali soil from the flowering stage to maturity stage and soil CAT activity at the maturity stage (41.16~48.82%). In normal soil, soil phosphatase and urease activities were enhanced by AMF at the flowering stage and the podding stage, respectively. AMF also increased the contents of soil organic matter, available phosphorus, and hydrolysable nitrogen, but decreased soil EC in saline-alkali soil. In addition to the significant increase in soil available phosphorus content, AMF had no significant effect on the physical and chemical properties of the soil and other soil nutrients in normal soil. AMF significantly increased pod biomass and yield in saline-alkali soil and normal soil, and improved their agronomic characteristics. In conclusion, peanut seeds coated with AMF improved the root morphological characteristics of peanuts and improved the physical and chemical properties in peanut rhizosphere, especially in saline-alkali soil. The process of rhizosphere soil nutrient transformation was also enhanced. Finally, AMF improved plant agronomic traits to increase the pod yield (16.5~21.9%). This study provides the theoretical basis and technical support for the application of AMF in peanut production in saline-alkali soil.

Molecular basis for inhibition of methane clathrate growth by a deep subsurface bacterial protein.

Methane clathrates on continental margins contain the largest stores of hydrocarbons on Earth, yet the role of biomolecules in clathrate formation and stability remains almost completely unknown. Here, we report new methane clathrate-binding proteins (CbpAs) of bacterial origin discovered in metagenomes from gas clathrate-bearing ocean sediments. CbpAs show similar suppression of methane clathrate growth as the commercial gas clathrate inhibitor polyvinylpyrrolidone and inhibit clathrate growth at lower concentrations than antifreeze proteins (AFPs) previously tested. Unlike AFPs, CbpAs are selective for clathrate over ice. CbpA3 adopts a nonglobular, extended structure with an exposed hydrophobic surface, and, unexpectedly, its TxxxAxxxAxx motif common to AFPs is buried and not involved in clathrate binding. Instead, simulations and mutagenesis suggest a bipartite interaction of CbpAs with methane clathrate, with the pyrrolidine ring of a highly conserved proline residue mediating binding by filling empty clathrate cages. The discovery that CbpAs exert such potent control on methane clathrate properties implies that biomolecules from native sediment bacteria may be important for clathrate stability and habitability.