Identification,Isolation and Genomic Characterization of Porcine Astrovirus in Shandong Province,China in 2021-2023

[Objective]The paper was to identify,isolate,and characterize porcine astrovirus in Shandong Province between 2021 and 2023.[Method]A total of 1025 samples of porcine diarrhea samples were collected from various regions of Shandong Province between January 2021 and October 2023.The samples were tested by RT-PCR,followed by sequencing and phylogenetic analyses of the polymerase.[Result]The total positive rate of PAstV was 34.6%(355/1025).The respective proportions of individuals infected with PAstV-1,PAstV-2,PAstV-4 and PAstV-5 were 25.4%(90/355),28.2%(100/355),35.2%(125/355)and 22.5%(80/355),respectively.Additionally,mixed infection was observed.Meanwhile,849 samples of healthy pigs were tested by RT-PCR,and the results demonstrated that the total positive rate of PAstV was 8.13%(69/849).Of these,the proportion of PAstV-1,PAstV-2 and PAstV-4 infection was 27.5%(19/69),37.7%(26/69)and 40.6%(28/69),and a mixed infection also existed.Further sequencing and characterization of some the selected isolates revealed low sequence identities(56.2%)with known PAstV strains,indicating the presence of novel types or genotypes of PAstVs.Furthermore,the isolation conditions of porcine astrovirus were optimized,resulting in the purification of a pure PAstV-4 strain(designated PAstV-4-GRF1).The virus was found to exhibit typical astroviral morphology,with nucleotide identity ranging from 89.9 to 95.4%with previously published PAstV-4 strains.Then,macrovirus transcriptome sequencing showed that 88.30%of the CRF1 samples were mammalian astroviruses.By species classification,PAstV 4 and PAstV 2 accounted for 21.79%and 0.32%,respectively.Phylogenetic tree analysis showed that the c15050 fragment was identical to the GRF-1 sequencing fragment of the isolated strain,and exhibited the highest homology with the Hunan PAstV-4 sequence MK460231 in China.[Conclusion]As the inaugural isolated PAstV-4 strain,it furnishes pivotal materialfor the investigation ofthe biological and pathogenic properties of this virus as well as for the prospectivedevelopment of relevant biological and diagnostic reagents.

Resolved concerns after 28 years of Bt cottonin China

Bacillus thuringiensis Berliner(Bt)cotton was widely grown in China from 1997.Since then,it has resulted in many misunderstandings and concerns about Bt cotton.However,extensive research and practical experience over the past 28 years in China have led to the resolution of many of these concerns.This short review explores how the concerns has been resolved,and provides valuable insights for the future utilization of genetically modified products.

The extraction of effective components and an antioxidant activity study of Tulipa edulis

Plant extracts from natural sources are an excellent choice for food additives and natural antioxidants.In this study,the active components of Tulipa edulis were extracted and analysed,and their antioxidant capacity was measured.Then,the crude extract mixture was separated and purified using a Sephadex LH-20 gel,and the antioxidant activity of the purified products was determined.Human umbilical vein endothelial human umbilical vein endothelial cells(HUVEC)cells were treated with 35 mmol/L glucose to construct a model of oxidative stress.Then,the cells were treated with the active component to observe whether the products of T.edulis could have a good protective effect on HUVEC cells induced by glucose.Transcriptome analysis was also performed on HUVEC cells after same treatment to explore the possible mechanism of the component F2 protecting HUVEC cells from oxidative stress induced by high glucose.The results showed that component F2 obtained from T.edulis has strong antioxidant activity.Moreover,F2 can play a strong antioxidant protective role in HUVEC cells.Meanwhile,the gene expression of heme oxygenase 1(HO-1),γ-glutamyl cysteine ligase catalytic subunit(GCLC)and NAD(P)H quinone oxidoreductase-1(NQO1)in HUVEC cells was up-regulated after treated with F2.This study provides reference value for the further development and application of T.edulis and the d evelopment of functional food.

Effects of water and nitrogen rate on grain-filling characteristics under high-low seedbed cultivation in winter wheat

A high-efficiency mode of high-low seedbed cultivation(HLSC)has been listed as the main agricultural technology to increase land utilization ratio and grain yield in Shandong Province,China.However,limited information is available on the optimized water and nitrogen management for yield formation,especially the grain-filling process,under HLSC mode.A three-year field experiment with four nitrogen rates and three irrigation rates of HLSC was conducted to reveal the response of grain-filling parameters,grain weight percentage of spike weight(GPS),spike moisture content(SMC),and winter wheat yield to water and nitrogen rates.The four nitrogen rates were N1(360 kg ha^(-1) pure N),N2(300 kg ha^(-1) pure N),N3(240 kg ha^(-1) pure N),and N4(180 kg ha^(-1) pure N),respectively,and the three irrigation quotas were W1(120 mm),W2(90 mm),and W3(60 mm),respectively.Results showed that the determinate growth function generally performed well in simulating the temporal dynamics of grain weight(0.989<R^(2)<0.999,where R2 is the determination coefficient).The occurrence time of maximum filling rate(T_(max))and active grain-filling period(AGP)increased with the increase in the water or nitrogen rate,whereas the average grain-filling rate(G_(mean))had a decreasing trend.The final 1,000-grain weight(FTGW)increased and then decreased with the increase in the nitrogen rates and increased with the increase in the irrigation rates.The GPS and SMC had a highly significant quadratic polynomial relationship with grain weight and days after anthesis.Nitrogen,irrigation,and year significantly affected the T_(max),AGP,G_(mean),and FTGW.Particularly,the AGP and FTGW were insignificantly different between high seedbed(HLSC-H)and low seedbed(HLSC-L)across the water and nitrogen levels.Moreover,the moderate water and nitrogen supply was more beneficial for grain yield,as well as for spike number and grain number per hectare.The principal component analysis indicated that combining 240-300 kg N ha^(-1) and 90^(-1)20 mm irrigation quota could improve grain-filling efficiency and yield for the HLSC-cultivated winter wheat.

Development of biofertilizers for sustainable agriculture over four decades(1980-2022)

The application of biofertilizers is becoming an inevitable trend to substitute chemical fertilizers for sustainable agriculture.To better understand the development of biofertilizers from 1980 to 2022,we used bibliometric mining to analyze 12,880 journal articles related to biofertilizer.The network cooccurrence analysis suggested that the biofertilizers research can be separated into three stages.The first stage(1980-2005)focused on nitrogen fixation.The second stage(2006-2015)concentrated on the mechanisms for increasing plant yield.The third stage(2016-2022)was the application of biofertilizers to improve the soil environment.The keyword analysis revealed the mechanisms of biofertilizers to improve plant-growth:biofertilizers can impact the nutritional status of plants,regulate plant hormones,and improve soil environments and the microbiome.The bacteria use as biofertilizers,included Pseudomonas,Azospirillum,and Bacillus,were also identified through bibliometric mining.These findings provide critical discernment to aid further study of biofertilizers for sustainable agriculture.

Mechanism of Tea Polyphenols in Alleviating Thermal Damage Based on Network Pharmacology

[Objective]This paper was to investigate the action targets and pathways of tea polyphenols in alleviating heat stress-induced injury by using network pharmacological analysis and an H9C2 cell model.[Method]First,the corresponding targets of tea polyphenols were obtained from the PubChem database.Then,the core targets were screened based on topological parameters.The relevant metabolism pathways of tea polyphenols related to diseases were identified through GO functional annotation and KECG signaling pathway enrichment.Moreover,common targets for thermal injury and targets of tea polyphenols were obtained.Then,GO functional annotation was performed to explore the pathway of tea polyphenols in alleviating heat stress damage.H9C2 cells were cultured at 42 C to construct the heat stress model,and the cells were treated with 10μg/mL tea polyphenols.The key genes were confirmed using RT-PCR technology.[Result]The study yielded 364 targets corresponding to tea polyphenols,including 68 core targets.These targets are related to various biological processes such as involve oxidative stress,cancer,lipopolysaccharide-mediated signaling pathways,antiviral responses,regulation of cellular response to heat,apoptosis,and cellular lipid metabolic metabolism.Tea polyphe nols alleviate thermal damage by targeting BCL2,HSP90AA1,HSPA1A,JUN,MAPK1,NFKB1,NFKBIA,NOS3,and TP53.Moreover,10 mg/L tea polyphenols were found to upregulate the transcription levels of Hsp70,HO-1,NQ-O1,Nrf2,and MAPKI,and the transcription levels of Bax/Bcl2,p38,and JNK were downregulated to alleviate the heat stress-induced injury.[Conclusion]Tea polyphenols may enhance the antioxidant ability of H9C2 cells and inhibit cell apoptosis,thereby reducing heat stress injury.

Analysis of Heavy Metal Sources for Vegetable Soils from Shandong Province, China

Heavy metal pollution in agricultural soils has serious negative influence on human health. Concentrations of Cd, Hg, As, Pb, Cr, Cu, Zn, and Ni in top soils （0-20 cm） of greenhouses and farmlands from four main vegetable production areas Shouguang, Laiyang, Jinxiang, and Zhangqiu in Shandong Province, one of the most rapidly developing regions in China, were measured in this study. Shouguang is mainly occupied by greenhouse vegetables and the other three areas are mainly open field culture. Total of 149 soil samples were collected. The average concentrations of the eight heavy metals of the tested 149 soil samples were all below the threshold values according to ＂Farmland environmental quality evaluation standards for edible agricultural products （HJ332-2006）＂ of China. However, most of the studied heavy metals were present at higher concentrations than those of the natural background levels in local agricultural soils. Among the total 149 soil samples, 22 samples were contaminated by Cd, Ni, Cu, or Hg. Comparisons showed that the main pollution element in greenhouse vegetable soils was Cd, while that of open field vegetable soils was Cu. The results of principal components analysis （PCA） suggested that concentrations of Cr, As, and Ni were mainly controlled by parent rocks; Hg and Pb were affected by anthropogenic activities such as vehicle and industrial fumes and waste water irrigation. Meanwhile, concentrations of Cd, Cu, and Zn were affected mainly by the use of agrochemicals. Most of the heavy metals were positively correlated with each other in concentration. Appropriate measures should be taken to effectively control heavy metal levels in vegetable soils and thus protect human health.

The Relationship Between Chinese Raw Dumpling Quality and Flour Characteristics of Shandong Winter Wheat Cultivars

Dumpling is one of the most important traditional wheat products in China. Dumpling quality is determined by the characteristics of both flour and filling, thus improvement of flour quality plays an important role in improving dumpling quality. Thirty-nine Shandong winter wheat cultivars and advanced lines sown in Jinan, Shandong Province, China, in the 2008-2009 cropping season were used to determine genetic variation in Chinese raw dumpling quality and its relationship with flour characteristics. Large variations were observed for protein quality parameters in comparison with starch properties. Variation in color of the raw dumpling sheet was broader than that of sensory evaluation parameters of boiled dumpling among tested wheat cultivars, indicating the large influence of filling on dumpling color. Two cultivars, Jimai 20 and Zimai 12, were identified as possessing very good quality of raw dumpling, and 21 cultivars and advanced lines showed good quality. Protein and total starch content influenced the L＊ value of raw dumpling sheets. L＂ value at 0 and 2 h after sheeting were significantly influenced by protein content （r=-0.46 and -0.52, P〈0.01） and total starch content （r=0.55 and 0.57, P〈0.01）, respectively. Flour yellow pigment was significantly corrected with a＊ （r=-0.67 and -0.62, P〈0.01） and b＊ （r=0.87 and 0.84, P〈0.01） value of raw dumpling sheets at 0 and 2 h after sheeting, respectively. Gluten strength parameters such as farinograph mixing tolerance index （MTI, r=-0.55, P〈0.01） were positively associated with appearance. MTI and energy were also significantly and positively correlated with elasticity of raw dumpling, with r=-0.54 and 0.47 （P〈0.01）. The positive relationships between peak viscosity （r=0.51, P〈0.01）, breakdown （r=0.54, P〈0.01）, and smoothness of raw dumpling were also observed. Therefore, it is suggested that breeding programs should give more attention to gluten strength and starch pasting parameters for raw dumpling quality improvement.

Status and Suggestions of the Pesticide Use in the Protected Vegetable Fields in Shandong Province

This paper briefly introduced the issues of food safety and environmental pollution caused by pesticide residues in protected vegetables, discussed the status and problems of pesticide use in the protected vegetables in Shandong Province, and analyzed the main factors leading to the pesticide residues, including the low education of most farmers, lack of correct identification of diseases and insect pests, use of pesticides based on personal experience, pesticide preparation by bare hands, large dose of pesticide, frequent application, pesticide spraying without protection, uneven spraying, leakage of pesticide from the sprayers, etc.. Finally, based on the vegetable planting features and advantages in Shandong Province, some suggestions were proposed for references, such as, to enhance the monitoring of pesticide residue, to improve the educational level of farmers and to scentifically use the pesticide.

Effects of Long-term Located Fertilization on Evolution of Available Phosphorus and Phosphorus Pool in Shandong Fluvo-aquic Soil

This study was conducted to investigate the effects of long-term located fertilization on soil phosphorus,the changes of soil available phosphorus(OlsenP),the evolution of soil total phosphorus(TP)and the ratio change of Olsen-P to TP(PAC)by 33-year fertilization experiments in winter wheat-summer maize rotation system in Shandong fluvo-aquic soil.Eight treatments were designed as no fertilization(CK),nitrogen fertilizer(N),nitrogen and phosphate fertilizer(NP),nitrogen and potassium fertilizer(NK),phosphate and potassium fertilizer(PK),nitrogen-phosphate-potassium fertilizer(NPK),reduced NPK fertilizer(N(15)PK),and increased NPK fertilizer(N_(25)PK).Meanwhile,eight organic fertilizer-added treatments were designed based on the application of inorganic fertilizer the same as the above ones.The results showed that TP,Olsen-P and PAC of treatments added with organic fertilizer were higher than those without organic fertilizer,and those of the treatments applied with phosphate fertilizer were higher than those of no phosphate fertilizer.With the increase of years,soil P pool decreased due to crop absorption,nutrient loss and morphological transformation and other causes under the treatments of without and only phosphate fertilizer,while remained stable under the treatments added with organic fertilizer.The PAC values were generally lower in fluvo-aquic soil,and it could be improved by the application of organic fertilizer.On the whole,the application of chemical phosphate fertilizer combined with organic fertilizer could improve the phosphorus content in soil and ensure the supply of phosphorus nutrition.This study would provide scientific basis for fertilization management and soil fertility in fluvo-aquic soil.

Identification,evolution,expression and protein interaction analysis of genes encoding B-box zinc-finger proteins in maize

The B-box(BBX)family of proteins consists of zinc-finger transcription factors with one or two highly conserved B-box motifs at their N-termini.BBX proteins play crucial roles in various aspects of plant growth and development,including seedling photomorphogenesis,shade avoidance,flowering time,and biotic and abiotic stress responses.Previous studies have identified many different BBXs from several plant species,although the BBX family members in maize are largely unknown.Genome-wide identification and comprehensive analysis of maize BBX(ZmBBX)expression and interaction networks would therefore provide valuable information for understanding their functions.In this study,36 maize BBXs in three major clades were identified.The ZmBBXs within a given clade were found to share similar domains,motifs,and genomic structures.Gene duplication analyses revealed that the expansion of BBX proteins in maize has mainly occurred by segmental duplication.The expression levels of ZmBBXs were analyzed in various organs and tissues,and under different abiotic stress conditions.Protein–protein interaction networks of ZmBBXs were established using bioinformatic tools and verified by bimolecular fluorescence complementation(BiFC)assays.Our findings can facilitate a greater understanding of the complexity of the ZmBBX family and provide novel clues for unravelling ZmBBX protein functions.

Inclusion of peanut in wheat–maize rotation increases wheat yield and net return and improves soil organic carbon pool by optimizing bacterial community

Improving soil quality while achieving higher productivity is the major challenge in the agricultural industry. Wheat(Triticum aestivum L.)–maize(Zea mays L.)(W–M) rotation is the dominant planting pattern in the Huang-HuaiHai Plain and is important for food security in China. However, the soil quality is deteriorating due to the W–M rotation’s long-term, intensive, and continuous cultivation. Introducing legumes into the W–M rotation system may be an effective way to improve soil quality. In this study, we aimed to verify this hypothesis by exploring efficient planting systems(wheat–peanut(Arachis hypogaea L.)(W–P) rotation and wheat rotated with maize and peanut intercropping(W–M/P)) to achieve higher agricultural production in the Huang-Huai-Hai Plain. Using traditional W–M rotation as the control, we evaluated crop productivity, net returns, soil microorganisms(SMs), and soil organic carbon(SOC) fractions for three consecutive years. The results indicated that wheat yields were significantly increased under W–P and W–M/P(382.5–579.0 and 179.8–513.1 kg ha-1, respectively) compared with W–M. W–P and W–M/P provided significantly higher net returns(58.2 and 70.4%, respectively) than W–M. W–M/P and W–M retained the SOC stock more efficiently than W–P, increasing by 25.46–31.03 and 14.47–27.64%, respectively, in the 0–20 cm soil layer. Compared with W–M, W–M/P improved labile carbon fractions;the sensitivity index of potentially mineralizable carbon, microbial biomass carbon(MBC), and dissolved organic carbon was 31.5, 96.5–157.2, and 17.8% in 20–40, 10–40, and 10–20 cm soil layers, respectively. The bacterial community composition and bacteria function were altered as per the soil depth and planting pattern. W–M/P and W–M exhibited similar bacterial community composition and function in 0–20 and 20–40 cm soil layers. Compared with W–P, a higher abundance of functional genes, namely, contains mobile elements and stress-tolerant, and a lower abundance of genes, namely,potentially pathogenic, were observed in the 10–20 cm soil layer of W–M and the 0–20 cm soil layer of W–M/P. SOC and MBC were the main factors affecting soil bacterial communities, positively correlated with Sphingomonadales and Gemmatimonadales and negatively correlated with Blastocatellales. Organic input was the main factor affecting SOC and SMs, which exhibited feedback effects on crop productivity. In summary, W–M/P improved productivity, net returns, and SOC pool compared with traditional W–M rotation systems, and it is recommended that plant–soil–microbial interactions be considered while designing high-yield cropping systems.

Effect of the L-D_(1)alleles on leaf morphology,canopy structure and photosynthetic productivity in upland cotton(Gossypium hirsutum L.)

One of the most important objectives for breeders is to develop high-yield cultivars.The increase in crop yield has met with bottlenecks after the first green revolution,and more recent efforts have been focusing on achieving high photosynthetic efficiency traits in order to enhance the yield.Leaf shape is a significant agronomic trait of upland cotton that affects plant and canopy architecture,yield,and other production attributes.The major leaf shape types,including normal,sub-okra,okra,and super-okra,with varying levels of lobe severity,are controlled by a multiple allelic series of the D-genome locus L-D_(1).To analyze the effects of L-D_(1)alleles on leaf morphology,photosynthetic related traits and yield of cotton,two sets of near isogenic lines(NILs)with different alleles were constructed in Lumianyan 22(LMY22)and Lumianyan 28(LMY28)backgrounds.The analysis of morphological parameters and the results of virus-induced gene silencing(VIGS)showed that the regulation of leaf shape by L-D_(1)alleles was similar to a gene-dosage effect.Compared with the normal leaf,deeper lobes of the sub-okra leaf improved plant canopy structure by decreasing the leaf area index(LAI)and increasing the light transmittance rate(LTR),and the mid-range LAI of sub-okra leaf also guaranteed the accumulation of cotton biomass.Although the chlorophyll content(SPAD)of sub-okra leaf was lower than those of the other two leaf shapes,the net photosynthetic rate(Pn)of sub-okra leaf was higher than those of okra leaf and normal leaf at most stages.Thus,the improvements in canopy structure,as well as photosynthetic and physiological characteristics,contributed to optimizing the light environment,thereby increasing the total biomass and yield in the lines with a sub-okra leaf shape.Our results suggest that the sub-okra leaf may have practical application in cultivating varieties,and could enhance sustainable and profitable cotton production.

Whole genome SNPs among 8 chicken breeds enable identification of genetic signatures that underlie breed features

Many different chicken breeds are found around the world,their features vary among them,and they are valuable resources.Currently,there is a huge lack of knowledge of the genetic determinants responsible for phenotypic and biochemical properties of these breeds of chickens.Understanding the underlying genetic mechanisms that explain across-breed variation can help breeders develop improved chicken breeds.The whole-genomes of 140 chickens from 7 Shandong native breeds and 20 introduced recessive white chickens from China were re-sequenced.Comparative population genomics based on autosomal single nucleotide polymorphisms(SNPs)revealed geographically based clusters among the chickens.Through genome-wide scans for selective sweeps,we identified thyroid stimulating hormone receptor(TSHR,reproductive traits,circadian rhythm),erythrocyte membrane protein band 4.1 like 1(EPB41L1,body size),and alkylglycerol monooxygenase(AGMO,aggressive behavior),as major candidate breed-specific determining genes in chickens.In addition,we used a machine learning classification model to predict chicken breeds based on the SNPs significantly associated with recourse characteristics,and the prediction accuracy was 92%,which can effectively achieve the breed identification of Laiwu Black chickens.We provide the first comprehensive genomic data of the Shandong indigenous chickens.Our analyses revealed phylogeographic patterns among the Shandong indigenous chickens and candidate genes that potentially contribute to breed-specific traits of the chickens.In addition,we developed a machine learning-based prediction model using SNP data to identify chicken breeds.The genetic basis of indigenous chicken breeds revealed in this study is useful to better understand the mechanisms underlying the resource characteristics of chicken.

Emergence of recombinant PCV2 between genotype 2a and 2b in pig herds in Shandong province of China

Since late 2005, the swine industry in Shandong province has experienced a significant increase in postweaning multisystemic wasting syndrome （PMWS）. Porcine circovirus type 2 （PCV2） is one of the major etiologic agents of PMWS. In order to understand the genetic diversity of PCV2 in swine herds in Shandong province, we sequenced six complete genomes of PCV2 between 2005 and 2007. Sequence analysis revealed that all the six PCV2 strains belonged to one genotype （named PCV 2b）. The deduced amino acid sequences of the replication associated protein （coded by ORFI） showed that all the six PCV2 strains belonged to genotype 2a and could be further divided into two clusters. Based on the analysis of the capsid protein （coded by ORF2）, the six PCV2 strains belonged to genotype 2b. All the six strains belonged to recombinant viruses between genotype PCV2a and genotype PCV2b. This study implied that such recombinant PCV2 spread widely in Shandong area and might be the reason that pig death rate associated with PMWS increased in recent years.

Residue behaviors and dietary risk of cyazofamid in turnip,onion and romaine lettuce assessed by a QuEChERS-LC-MS/MS methods

In this study, the dissipation, terminal residues and dietary risk of the cyazofamid residue in three globally consumed minor crops, turnip(Brassica rapa L.), onion(Allium cepa L.)and romaine lettuce(Lactuca sativa L.)were assessed by supervised field trials at 6 locations of China in 2020. A reliable QuEChERS-LC-MS/MS method was developed for simultaneous analysis of cyazofamid and its primary metabolite, 4-chloro-5-p-tolylimidazole-2-carbonitrile(CCIM). The average recoveries of cyazofamid and CCIM in turnip, onion and romaine lettuce were 79%–97% and 77%–99%, respectively. The half-lives of cyazofamid in these crops were 5.7–6.5, 5.3–8.7 and 5.8–6.5 days in turnip, onion and romaine lettuce, respectively, with the final cyazofamid residues at harvest all lower than limit of quantitation(LOQ, 0.01 mg/kg). The turnip roots, onion bulbs and romaine lettuce leaves grown in China under Good Agricultural Practices conditions and harvested 10, 14 and 5 days, respectively, after the cyazofamid SC(100 g/L)application, would be reasonably safe for consumption by the general population. The maximum residue limit(MRL)values of cyazofamid could be tentatively set as 10 mg/kg for turnip leaves, 0.3 mg/kg for turnip roots, 0.05 mg/kg for onion bulbs, and 10 mg/kg for romaine lettuce leaves.

Observation of Flowering Process of Grape(Vitis vinifera L.)—Insight into the Starting of Pollination of Flower Hiding in Calyptras(Cap)

It is generally agreed that many Vitis vinifera L.cultivars are self-fertile,where self-pollination often occurs before capfall in a process called cleistogamy.Therefore,it is difficult to identify the right time to remove stamens before self-pollination during the cross-breeding of grape.For this paper,we observed the process of grape flowering and measured the pollen viability and stigma receptivity of grape flowers of‘Shine Muscat’in order to identify the starting time of self-pollination before capfall and to provide useful information for improving the efficiency of cross-breeding.The results demonstrate that the anther is not cracked during the visible clusters and separated clusters stages.Meanwhile,in the separated floral buds,flowering begins,and full bloom stages,the pollen viability is 60.7%,73.2%and 80.3%,respectively;however,at the berry set stage,pollen viability drops to zero.The top of the mature stigma is composed of a layer of nearly cylindrical papillary cells,and the stigma receptivity for pollen changes with the development of flowers:in particular,no reaction was observed in the visible clusters stage;weak positive reaction at the separated clusters stage;strong positive reaction at the separated floral buds,flowering begins,and full bloom stages;and no reaction at the berry set stage.In the separated floral buds stage,pollen tubes were seen germinating in the style.In the flowering begins stage,more pollen tubes were observed at the entry of the ovary.During the full bloom stage,most pollen tubes elongated into the ovary base and some entered the pearl hole.At the berry set stage,newborn endosperm nucleus could be seen in the ovule.From the above,we can conclude that the initiation time of closed fertilization for‘Shine Muscat’grape can be judged as the separated floral buds stage,and it is best to discard the stamen before the separated floral buds stage when conducting cross-breeding.

Personal glucose meters coupled with signal amplification technologies for quantitative detection of non-glucose targets:Recent progress and challenges in food safety hazards analysis

Ensuring food safety is paramount worldwide.Developing effective detection methods to ensure food safety can be challenging owing to trace hazards,long detection time,and resource-poor sites,in addition to the matrix effects of food.Personal glucose meter(PGM),a classic point-of-care testing device,possesses unique application advantages,demonstrating promise in food safety.Currently,many studies have used PGM-based biosensors and signal amplification technologies to achieve sensitive and specific detection of food hazards.Signal amplification technologies have the potential to greatly improve the analytical performance and integration of PGMs with biosensors,which is crucial for solving the challenges associated with the use of PGMs for food safety analysis.This review introduces the basic detection principle of a PGM-based sensing strategy,which consists of three key factors:target recognition,signal transduction,and signal output.Representative studies of existing PGM-based sensing strategies combined with various signal amplification technologies(nanomaterial-loaded multienzyme labeling,nucleic acid reaction,DNAzyme catalysis,responsive nanomaterial encapsulation,and others)in the field of food safety detection are reviewed.Future perspectives and potential opportunities and challenges associated with PGMs in the field of food safety are discussed.Despite the need for complex sample preparation and the lack of standardization in the field,using PGMs in combination with signal amplification technology shows promise as a rapid and cost-effective method for food safety hazard analysis.

Genome-wide association study for numbers of vertebrae in Dezhou donkey population reveals new candidate genes

Numbers of vertebrae is an important economic trait associated with body size and meat productivity in animals.However,the genetic basis of vertebrae number in donkey remains to be well understood.The aim of this study was to identify candidate genes affecting the number of thoracic(TVn)and the number of lumbar vertebrae(LVn)in Dezhou donkey.A genome-wide association study was conducted using whole genome sequence data imputed from low-coverage genome sequencing.For TVn,we identified 38 genome-wide significant and 64 suggestive SNPs,which relate to 7 genes(NLGN1,DCC,SLC26A7,TOX,WNT7A,LOC123286078,and LOC123280142).For LVn,we identified 9 genome-wide significant and 38 suggestive SNPs,which relate to 8 genes(GABBR2,FBXO4,LOC123277146,LOC123277359,BMP7,B3GAT1,EML2,and LRP5).The genes involve in the Wnt and TGF-βsignaling pathways and may play an important role in embryonic development or bone formation and could be good candidate genes for TVn and LVn.

MdAIL5 overexpression promotes apple adventitious shoot regeneration by regulating hormone signaling and activating the expression of shoot development-related genes

Adventitious shoot(AS)regeneration is a significant factor in the genetic transformation of horticultural plants.It is also a noteworthy approach to their vegetative propagation.AS regeneration remains highly dependent on the genotype or maturity of explants.We here found that the AS regeneration abilities of apple leaves were positively correlated with MdAIL5 expression.MdAIL5 overexpression dramatically increased AS regeneration efficiency.Notably,MdAIL5 overexpression could restore the AS formation ability of explants to a certain extent,which was lost with an increase in maturity.Endogenous hormone detection revealed that MdAIL5 overexpression changed the contents of auxin,cytokinin(CK),and other hormones in apple leaves.Transcriptome analysis revealed that many genes related to auxin,CK,and brassinolide signaling pathways were significantly and differentially expressed between MdAIL5-overexpressing transgenic apple and wild-type apple plants.Yeast one-hybrid assays,the electrophoretic mobility shift assay,and the dual-luciferase reporter assay revealed that MdAIL5 directly binds to MdARF9 and MdHB14 promoters and positively affects their expression.We here established a model of MdAIL5 regulating AS formation,which acts as a theoretical basis for facilitating genotype-or explant maturity-independent AS regeneration in the future.