Bioinformatics and expression patterns of Dicer-like(DCL),Argonaute(AGO)and RNA-dependent RNA polymerase(RDR)gene families in the whole genome of Solanum habrochaites were analyzed,so as to provide references for further study on the functions of DCL,AGO and RDR gene families in the response of S.habrochaites to abiotic and viral infection.Using Arabidopsis thaliana DCL,AGO and RDR genes as reference sequences,the genome of S.habrochaites LA1777 was searched by local perl language and software such as Pfam and SMART,and the members of ShDCL,ShAGO and ShRDR gene families were determined.Bioinformatics analysis of DCL,AGO and RDR family genes in S.habrochaites was carried out by means of ExPASy,GSDS 2.0,MEGA,Tbtools and SWISS-MODEL.According to abiotic stress treatment,Tomato chlorosis virus(ToCV)treatment and Real-time Fluorescence Quantitative PCR technology,the expression patterns of these genes were analyzed.Seven ShDCL,15 ShAGO and 6 ShRDR genes were identified from S.habrochaites,which were distributed on chromosome 5,7 and 6 respectively.The encoded proteins were similar in structure to DCL,AGO and RDR in other plants,and all of them contained conserved domains unique to this family.Phylogenetic analysis showed that these genes were divided into 4 subgroups,and there were high structural and functional similarities between S.habrochaites and S.lycopersicum.ShDCL2a,ShDCL2c,ShDCL3,ShDCL4,ShAGO1b,ShAGO3,ShAGO4b,ShAGO5,ShAGO7,ShAGO10a,ShAGO10b,ShRDR1,ShRDR2,ShRDR3a,ShRDR6a and ShRDR6b were significantly up-regulated after various abiotic stresses and ToCV infection.It is speculated that these genes play important roles in abiotic stress and virus infection.

In order to study the problem of strong winter/spring Brassica napus seed germination and flowering period under different winter sowing dates.Two strong winter rapeseeds and two spring rapeseeds provided by Gansu Agricultural University were used as materials.The experiment was carried out in the experimental field of Gansu Agricultural University from October 2022 to August 2023.The winter rapeseeds was carried out on October 11,2022.The winter/spring rapeseeds was sown every 20 days from December 10,2022,and the sowing ended on February 8,2023.The flowering period was recorded,and the germination seeds of winter rapeseed were sampled every 20 days to determine their physiological and biochemical characteristics and analyze the expression characteristics of vernalization genes(FLC,VRN2,FRI,FT).The results showed that the flowering period of winter/spring rape seeds was different by 22—34 days.The difference of flowering time between autumn sowing and spring sowing was 4—7 days.The flowering time of winter rapeseed in autumn sowing(October 11 th)was close to that of spring rapeseed under different winter sowing dates(December 10th,December 30th,January 19th,February 8th),and the flowering overlap time was as long as 15—20 days.With the delay of the sowing date,the relative expression levels of FLC,FRI and FT genes in germinating seeds of winter sowing were down-regulated.The relative expression of VRN2 gene was down-regulated in the early vernalization and up-regulated in the late vernalization.The activities of superoxide dismutase(SOD),peroxidase(POD),catalase(CAT)and the contents of soluble protein(SP),gibberellin(GA3)and salicylic acid(SA)in germinating seeds were increased in the early vernalization,but those were decreased in the late vernalization.The contents of malondialdehyde(MDA)and abscisic acid(ABA)were increased in rapeseed germinating with the increase of vernalization time.

This study explored the effects of intercropping and rotation on the growth,yield and quality of continuous cropping peanut,to provide theoretical basis for achieving high yield in peanut production.From 2022 to 2023,sweet potato-peanut rotation system(PSP)and maize-peanut intercropping and rotation system(PMP)were set up in the experimental farm of Henan University of Science and Technology on the basis of continuous cropping peanut for 2 years and 11 years respectively,with continuous cropping peanut as control(CCP1 and CCP2,respectively).The effects of PSP and PMP on photosynthetic characteristics,root characteristics,dry matter accumulation and distribution and yield of peanut were studied.The results showed that compared with CCP1,the leaf area index(LAI)of rotating peanut in PSP system(SRP)was significantly increased by 35.08%—53.68% and 24.32%—33.52% at pod-setting stage(PSS)and full pod maturity stage(PMS),respectively.The SPAD value at PSS and pod bulking stage(PBS)increased by 11.93%—18.55% and 5.95%—9.63%,respectively.Compared with CCP2,the LAI of rotating peanut in PMP system(MRP)increased by 46.81%—57.96% and 27.00%—61.78% at PSS and PMS,respectively.At PSS and PBS,compared with CCP2,the SPAD value of MRP and intercropping peanut(MIP)increased by 3.32%—3.69%,7.50%—8.64% and 5.47%—18.37%,15.73%—31.11%,respectively.At PSS and PBS,compared with CCP1,the net photosynthetic rate of SRP increased by 23.68%—41.31% and 26.52%—32.55%,and compared with CCP2,MRP increased by 12.77%—17.81% and 16.88%—62.07%,respectively.They both significantly improved the root length and root tip number,and promoted the dry matter accumulation and the distribution to pods during PMS,and the yields increased by 31.42%—47.36% and 54.12%—75.09%,respectively.Compared with CCP2,MIP reduced the LAI,net photosynthetic rate,root length,root tip number,as well as dry matter accumulation and yield of peanut under the influence of maize shading.At the same time,the content of peanut oleic acid and oleic acid-linoleic acid ratio was significantly increased after rotation.Among them,SRP increased by 1.63—1.65 percentage point and 6.59%—10.52%,respectively,compared with CCP1,and MRP increased by 1.95—2.82 percentage point and 9.75%—14.16% compared with CCP2,respectively.In summary,sweet potato-peanut rotation and maize-peanut rotation increased the peanut yield compared with continuous cropping peanut,the reason was that sweet potato-peanut and maize-peanut rotation promoted peanut root growth,delayed the leaf senescence,and increased photosynthetic rate,especially the photosynthetic rate during late growth period,which promoted the dry matter accumulation and distribution to seeds.Besides that,they could improve the quality of peanut to a certain extent.

This study systematically investigates the effects of silicon-modified biochar (MSC) on the chemical properties of acidic soil,organic carbon and silicon fractions,and the growth of tomato plants.Silicon-modified biochar was prepared,with a focus on investigating its impacts on carbon and silicon chemical fractions,and the availability in acidic soils;tomato growth and soil microbial activity were also evaluated.The results showed that silicon-modified biochar significantly increased soil pH,cation exchange capacity,electrical conductivity,available phosphorus and potassium.MSC also raised the levels of water-soluble sodium and iron in the soil and enhanced the activities of hydrogen peroxidase and sucrase enzymes,thereby improving soil quality.Both biochar modification and unmodified biochar significantly increased the content of different carbon fractions in the soil.Compared with unmodified biochar,silicon-modified biochar significantly increased soil microbial biomass carbon(21.9%) and water-soluble organic carbon (898.3%).Furthermore,silicon-modified biochar significantly increased the contents of soil available silicon,water-soluble silicon,free silicon,active silicon,iron-manganese-bound silicon and amorphous silicon by 362.6%,158.9%,18.1%,34.9%,193.8%,and 74.1%,respectively.Meanwhile,the application of biochar promoted the growth of tomato plants and the absorption of silicon nutrients,with modified biochar showing more pronounced effects.The accumulation of plant dry matter,silicon content,and absorption rate increased by 82.0%,98.9%,and 261.5%,respectively.In summary,silicon-modified biochar significantly affected the carbon and silicon chemical forms and transformation in the soil,increased soil effectiveness and enzyme activity,thereby promoting nutrient absorption and growth of crops,demonstrating its good potential application in agricultural production.

To study the photosynthetic characteristics,fluorescence parameters and yield response to phosphorus in maize,and to clarify the optimal phosphorus application rate for maize under drip irrigation and water fertilization technology.Providing solid theoretical basis and technical support for high-yield and high-efficiency cultivation of maize in Ningxia region.The experiment was carried out at Pingjipu Farm,Yinchuan,Ningxia,from 2019 to 2020,with six phosphorus treatments in the order of 0(P0),60(P1),120(P2),180(P3),240(P4),and 300 kg/ha(P5).Analysis of the changing patterns of photosynthetic and fluorescence parameters of spring maize leaves and their correlation with yield under different phosphorus fertilizer treatments.In two years,during the big bell mouth stage,leaf area index (LAI) was increased by 4.21% to 12.78% and 4.68% to 15.60% for P3 compared to other treatments,respectively.Phosphorus fertilizer at 180 kg/ha was most effective in promoting leaf area index and photosynthetic potential(LAD) of maize.LAD was significantly increased by 14.42% under P3 treatment compared to no phosphorus fertilizer treatment during the full two year period.The photosynthetic characteristics of maize responded differently to the intensity of phosphorus application,and as the intensity of phosphorus application increased,the net photosynthetic rate (Pn) all reached the maximum value after the stamen pumping stage,and at the R1 stage of the 2 years,the Pn was significantly increased by 10.68% under the P3 treatment as compared to the no-phosphorus-fertilizer treatment.The coefficient of determination (R²) was 0.926 5,0.889 9,and 0.832 0,respectively.Phosphorus application increased the maize photosystem Ⅱ composite performance index (PI),which had its maximum peak at the R1 stage in 2 years,and PI increased by 1.12% to 8.50% and 8.47% to 15.40% under the P3 treatment compared with the other treatments,respectively.The maximum yield was obtained at 180 kg/ha of phosphorus application,which was 17.27% higher as compared to no phosphorus treatment.Based on the analysis of the yield fitting equation,it was shown that the maximum corn yield of 13 823.84 kg/ha was reached at 179.34 kg/ha of phosphorus applied.Pearson's correlation analysis showed that appropriate leaf area index significantly affected maize yield in the late stage,and the photosynthetic parameters all had highly significant effects on maize yield completion;principal component analysis showed that the P3 treatment had the highest composite score for the optimization effect on maize yield.Reasonable transportation of phosphorus fertilizer can effectively ensure higher SPAD value,PSⅡ reaction center activity,improve the capture and utilization of light energy in spring maize,and promote photosynthesis,so as to improve the yield and economic benefits of maize.

In order to clarify the role of the Zn(Ⅱ)2Cys6 transcription factor gene VDAG_ 04814 in the growth, development and pathogenicity of the Verticillium dahliae. It constructed a VDAG_04814 gene knockout mutant using homologous recombination mediated by polyethylene glycol. Wild-type and mutant strains were inoculated separately onto PDA media supplemented with hydrogen peroxide, sodium chloride, potassium chloride, sorbitol, sodium dodecyl sulfate, Congo red, as well as onto media overlaid with sterile cellophane, to analyze their levels of resistance to oxidative stress, salt stress, osmotic stress, stress on cell wall and plasma membrane integrity, and strain penetration ability. Their pathogenicity was assayed, and the fungal biomass in potato plants was detected. After hygromycin selection and PCR validation, the correct knockout transformants were able to amplify DNA bands of 1 500 bp upstream and downstream, respectively, as well as the full-length 4 500 bp knockout fragment sequence. The results demonstrated that the growth rate and melanin formation ability of the ΔVDAG_04814 mutants were significantly reduced. On media subjected to oxidative stress and salt stress with the addition of hydrogen peroxide, sodium chloride, and potassium chloride, ΔVDAG_04814 showed a higher inhibition rate compared to the wild-type. On osmotic stress media with sorbitol, the growth inhibition rate of ΔVDAG_04814 was significantly lower than the wild type. No growth inhibition was observed for ΔVDAG_04814 on media subjected to cell wall and membrane integrity stress with the addition of sodium dodecyl sulfate and Congo red. On media overlaid with sterile cellophane, no colonies grew for ΔVDAG_04814, whereas the wild-type strain produced normal colonies. Pathogenicity tests indicated that the wilting index of ΔVDAG_04814 was significantly reduced compared to the wild-type, with wilting index ranging from 47.22 to 55.56. It has been demonstrated that VDAG_04814 can regulate the growth, development, stress resistance, penetration ability and pathogenicity of V. dahliae towards potato. This study provides a new target for the control of potato Verticillium wilt disease.

Investigating the predominant genotypes of Bovine viral diarrhea virus(BVDV)infecting cattle in Tongliao,Inner Mongolia,to provide reference for the BVDV epidemiology and prevention and control.In the preliminary phase of the experiment,fecal samples from diarrheic calves were collected at a cattle farm in Tongliao,Inner Mongolia.These samples were tested using PCR to detect BVDV positivity.Positive fecal samples were then inoculated into madin-darby bovine kidney cells(MDBK)for isolation.The isolated strains were identified using RT-PCR and indirect immunofluorescence staining.Subsequently,the full-length genome of the isolates was sequenced,followed by genetic evolution analysis and genotype determination based on sequences of the 5'UTR,N^pro,and E2 genes.The results indicated that this experiment successfully isolated a strain of BVDV,designated as NM-21.Inoculation of NM-21 into MDBK did not induce cytopathic effects,indicating it was a non-cytopathic strain(NCP).Both RT-PCR and indirect immunofluorescence staining confirmed its positivity,with a virus titer of 10^-3 TCID₅₀/mL.Based on the full-length genomic sequence,and homology and genetic evolution analysis of the 5'UTR,N^pro,and E2 gene sequences,the isolate NM-21 showed the highest nucleotide homology with the BVDV-1c subtype strain NM2103(GenBank accession number ON337882.1)from Inner Mongolia,China.

The highly contagious gastrointestinal infectious disease caused by Porcine epidemic diarrhea virus(PEDV)has led to significant economic losses in China's pig industry.It aimed to establish a basis for PEDV antibody detection methods and functional research of the N protein through the screening of specific nanobodies(Nbs)using phage display technology.Peripheral blood lymphocytes were isolated from a camel immunized with the N protein,and total RNA was extracted and reverse transcribed into cDNA.The variable domain of the heavy chain of heavy chain antibodies(VHH)was amplified by PCR,subcloned into the pCANTAB5E-ccdb vector,and electroporated into ER2738 competent cells to construct the VHH phage antibody display library.Subsequently,the library was subjected to four rounds of panning against the PEDV N protein,and positive phage clones were cloned into the pET-30a vector.The binding affinity and specificity of the Nbs were determined by indirect ELISA and Western Blot.The results showed that after the fifth immunization,the antibody titer reached 1∶25 600.The constructed phage display library had a capacity of 4.72×10⁸ and an abundance of 4.3×10¹⁰ cfu/mL,with a 93.75% positive rate.After four rounds of screening,16 Nb clones with different amino acid sequences were obtained,and Nb45 was validated to possess excellent specificity and binding ability to the PEDV N protein.This study successfully screened and obtained specific N protein-targeting Nbs,providing biological materials for the establishment of PEDV detection methods and foundational research.

In order to investigate the function of watermelon calcium-dependent protein kinase (CDPK) in grafted seedlings and abiotic stress environments, this study used RT-PCR technology to clone the ClCDPK(Cla97C01G019720) gene from watermelon grafted seedlings and performed bioinformatics analysis on it. Further designed specific primers with Kpn Ⅰ and Sal Ⅰ enzyme cleavage sites based on the ClCDPK sequence,conducted amplification and double enzyme cleavage, and connected with pCAMBIA1300 to successfully construct the expression vector pCAMBIA1300-35S-ClCDPK for the target gene.Using RT-qPCR technology, the gene expression levels of ClCDPK were measured in self rooted seedlings (ZG) and grafted seedlings (JJ) after being subjected to salt and drought stress, respectively.The results showed that the ORF of ClCDPK gene was 1 647 bp, encoding 548 amino acids. Its protein contained STKc_CAMK and FRQ1 functional domains, and was a hydrophilic protein. Subcellular localization prediction showed that the protein was located in the nucleus. Evolutionary tree analysis of ClCDPK with CDPK from six other plants revealed that it was closely related to CDPK from Cucurbitaceae melons and pumpkins, with protein sequence homology alignment exceeding 92.64%, indicating high homology.The RT-qPCR expression results showed that the expression level of ClCDPK in grafted seedlings was significantly higher than that in self rooted seedlings. With the duration of stress, the expression levels of ClCDPK in grafted and self rooted seedlings first increased and then decreased, and under the same stress treatment, the expression level of ClCDPK in grafted seedlings was higher than that in self rooted seedlings.This study indicated that ClCDPK responded positively to salt and drought stress, and the ability of grafted seedlings to resist stress was higher than that of self rooted seedlings. It is speculated that ClCDPK is one of the key factors in watermelon's response to grafting, thereby improving the salt and drought resistance of watermelon grafted seedlings.

To investigate the role of calcium-dependent protein kinase (CDPK) in wheat growth and stress response,the TaCDPK17 gene was cloned from common wheat and its sequence structure,expression pattern,and stress resistance function were preliminarily analyzed.The results showed that the length of the TaCDPK17 gene coding region was 1 701 bp, encoding 566 amino acids and possessing typical structural features of the CDPK family, including one conserved serine/threonine kinase domain and four EF hand shaped domains. Evolutionary tree analysis of TaCDPK17 and CDPK17 from 12 other plants showed that TaCDPK17 had high homology with the CDPK17 sequence of gramineous crops,especially Aegilops tauschii and barley.The promoter region of TaCDPK17 gene contained multiple cis regulatory elements related to hormone signaling pathways,light response.Among them, there are more abscisic acid (ABA) responsive elements (ABRE) and methyl jasmonate responsive elements (CGTCA). The expression analysis based on Real-time Fluorescence Quantitative PCR showed that the expression level of TaCDPK17 increased to varying degrees after induced by 100 μmol/L ABA, 100 μmol/L methyl jasmonate, 20% PEG6000, and 250 mmol/L NaCl. Under stress conditions of 2 μmol/L ABA and 100 mmol/L NaCl, the germination rate of Arabidopsis seeds overexpressing TaCDPK17 was significantly higher than that of the wild type. Meanwhile, overexpression of TaCDPK17 alleviated the inhibitory effects of ABA or osmotic stress treatments on seedling root growth. During stomatal closure, transgenic plants overexpressing TaCDPK17 are more sensitive to ABA and exhibit a stronger stomatal closure trend compared to wild-type plants. These results indicated that TaCDPK17 plays an important role in stress response and hormone signaling in wheat.

In order to explore the effects of different irrigation methods and nitrogen fertilizer management on late rice growth,development,yield formation,and nitrogen use efficiency,Y Liangyou 911 was used as test material.Two irrigation methods(W1.flooded irrigation;W2.moistening irrigation) and three nitrogen fertilizer managenments base,tiller,ear and grain fertilizer ratios(N1:5∶3∶2,N2:3∶4∶3 and N3:3∶4∶2∶1) were designed.No fertilization served as the control (CK1.flooded irrigation;CK2.moistening irrigation).Leaf area index,SPAD value of rice leaves,dry matter mass,yield formation and nitrogen use efficiency were determined under each treatment combination.The results showed that compared with W1,the LAI of rice treated with W2 was lower in the early growth stage and higher in the middle and late growth stages.The SPAD value of W1 treatment was higher,but there was no significant difference in SPAD value in the late growth period.Under the same irrigation conditions,compared with N1,N2 and N3 treatments it could delay the decline of LAI and SPAD values in the late growth period of rice.W2 treatment could significantly increase rice dry matter accumulation by 6.61%-16.37% compared with W1 treatment.Under nitrogen fertilizer treatment,the dry matter mass was higher in the early and middle stages of growth with N1,and the dry matter mass was higher in the later stages with N1 and N3.The yield increase of W2 mode was 7.59%-10.47% compared with W1 mode.The yield of W2N3 treatment was 3.24%-14.53% higher than that of other treatment.Although the effective panicle number was lower in W2N3 treatment,other yield components were increased,which resulted in higher yield.During the two years,W2N2 and W2N3 had higher values of total nitrogen accumulation and nitrogen absorption efficiency,W2N3 had higher values of nitrogen agronomic utilization,nitrogen partial productivity and nitrogen harvest index,and W1N3 had the higher value of physiological nitrogen utilization.In conclusion,irrigation methods and nitrogen management significantly affect rice yield and nitrogen uptake and utilization.The W2 (moistening irrigation) coupled with N3 (base fertilizer∶tillering fertilizer∶ear fertilizer∶grain fertilizer=3∶4∶2∶1) nitrogen management method is more conducive to rice dry matter accumulation,yield improvement and efficient use of nitrogen fertilizer,which can not only meet high yield,but also play a role in water saving.It is the best coupling method of water and fertilizer.

In order to investigate the effects of density on individual and population structure characteristics and yield of winter wheat under rainfed and limited water supply conditions,a field experiment was carried out at Gaocheng Experimental Station,Shijiazhuang City during 2022-2023 season using JM22 wheat cultivar under four densities:which were 1.8×10⁶ (D180),3.0×10⁶ (D300),4.2×10⁶ (D420) and 5.4×10⁶ (D540) plants/ha.Two irrigation treatments for each density,which were no irrigation during the whole growth period (W0) and irrigated once at the jointing stage (W1).The influence of planting density and irrigation treatments on leaf area (flag leaf,top 2 nd leaf,top 3 nd leaf,top 4 th leaf),non-leaf green organs (ear,awns,stem sheath) area,leaf area index,non-leaf green organ area index,dry matter accumulation,photosynthetic active radiation interception rate,water use efficiency (WUE) and yield of winter wheat were studied.The results showed that the leaf area of each leaf layer and non-leaf green organs decreased with the increase of density.D300 treatment got the highest leaf area index and non-leaf green organs index,and significantly higher than that in D540.The contribution of post-anthesis dry matter to grain yield was more than 70%.Compared with D540 treatment,reducing plant density decreased the transfer of dry matter before anthesis,but increased the accumulation of dry matter after anthesis and its contribution to grain yield.With the increase of density,WUE increased first and then decreased,D300 treatment achieved the highest WUE.Specifically,the WUE of D300 was 1.2%-14.4% and 2.5%-12.7% higher than that of other densities under W0 and W1,respectively.Compared with W0,W1 treatment increased the area of leaf and non-leaf green organs of different densities,delayed leaf senescence,and improved the photosynthetic active radiation interception rate of canopy.Ultimately,the grain yield increased by 28.1%-39.7%.Under the conditions of this experiment,D300 treatment increased the leaf and non-leaf green organ area,leaf and non-leaf green organ area index,canopy photosynthetically active radiation interception rate,post-anthesis dry matter accumulation and its contribution to final grain yield.The yield of D300 was 2.4%-6.6% and 0.3%-9.7% higher than that of the other densities under W0 and W1,respectively,which was the optimal density in this study.

The adult plant resistance gene Lr12 exhibits excellent resistance in production systems.To fine map and develop reliable molecular markers for Lr12,a cross was made between the susceptible variety Thatcher and the resistant near-isogenic line RL6011 containing the Lr12 gene.The F₁ generation resulting from this cross was self-pollinated to generate F₂ individual plants and F_2∶3 families.Field evaluations were conducted using a mixture of five highly virulent leaf rust pathotypes (PHTT, THKS, THTT, PHTS, and PHKS) to inoculate F₂ individual plants and F_2∶3 families for adult plant resistance assessment and genetic analysis of resistance.Subsequently,genotyping was performed using a 16K liquid chip on 10 resistant and 10 susceptible individuals from the F₂ generation to identify SNP markers closely linked to Lr12.This enabled the determination of the chromosomal physical interval containing the resistance gene,the development of SSR molecular markers,and the construction of a genetic linkage map.The results indicate that the segregation ratio of resistance to leaf rust in 3 494 F₂ individuals derived from the RL6011(Lr12)/Thatcher cross was consistent with a 3∶1 ratio ( {\mathrm{\chi }}_{3:1}^2=0.14;P=0.71). In the assessment of 685 F_2∶3 families, the segregation ratio among resistant individuals, resistant heterozygous individuals, and susceptible individuals conformed to a 1∶2∶1 ratio ( {\mathrm{\chi }}_{1:2:1}^2=2.01;P=0.37), suggesting that Lr12 is a dominant gene and the population segregation follows Mendelian single-gene inheritance patterns. Genetic linkage map analysis localized the adult plant leaf rust resistance gene Lr12 between SSR molecular markers YK12817 and YK12928,within a genetic interval of 0.38 cM.This corresponds to a physical interval of 2.09 Mb within the physical range of 579.44 Mb to 581.53 Mb on chromosome 4BL of the Chinese Spring reference genome(IWGSC.Ref.V1.0).These findings provide a solid basis for predicting candidate genes.

In order to analyze the possible role of dTLR2 in duck innate immunity,we study the expression of dTLR2 mRNA in immune organs during 1 to 10 weeks,as well as the expression in blood after challenging with Escherichia coli and Salmonella enteritis by Fluorescence Quantitative PCR.We also prepared polyclonal antibodies of the extracellular domain of dTLR2 by recombinant protein expression for further studying the biological function of dTLR2.The results showed that dTLR2 mRNA was expressed in spleen,thymus and bursa at different weeks.The expression of dTLR2 mRNA was significantly different among different weeks in the spleen and thymus tissues,and there was no significant difference in bursa.After infection with E.coli and S.enteritis,the expression of dTLR2 mRNA was significantly higher than that of the control group in the first and second days,but there was no significant difference among groups in the third day.Analyzing the sequence of dTLR2 gene,we predicted the extracellular domain,designed primers for PCR assay,and built pET28a-TLR2 recombinant plasmid.The recombinant plasmid was translated into E.coli BL21 (DE3) for expression of dTLR2-His protein.The SDS-PAGE results showed that the fusion recombinant protein efficiently expressed in the supernatant,molecular mass was around 29 ku.The results of Western Blot showed that the antibody which harvested from immunizing rabbits could detect recombinant dTLR2 and endogenous dTLR2.The analysis of the expression of dTLR2 and successful preparation of polyclonal antibody provide favorable support for further studying of dTLR2 biology function.

This study aimed to detect the SNPs of CDKN2A gene in Wenshang Barred chicken,and perform the analysis of its bioinformatics and expressions in skin tissues.Based on the high quality chromosome-level genome of Wenshang Barred chicken assembled by the research group of Resource Conservation and Evaluation in Jiangsu Institute of Poultry Science,the sequences were blasted to chicken reference genome (GRCg7b) and the SNPs were acquired.These SNPs were detected and genotyped by MassARRAY nucleic acid mass spectrometry in the chicken population.Bioinformatics software was used to predict the regulatory sequence of CDKN2A gene.The physicochemical properties and structural characteristics of the encoded protein were analyzed.The expressions of this gene in the skin tissues of Wenshang Barred chicken at different periods were detected by transcriptome and Real-Time Fluorescence Quantitative PCR.The results showed that the total length of CDKN2A and its CDS were respectively 9.854 kb and 183 bp,encoding 60 amino acids.Forty-one SNPs were found in this gene and its upstream and downstream 1 kb,of which four SNPs were newly discovered loci.These four new SNPs were located in the downstream and the intron,respectively.The first exon had two SNPs,both of which were missense mutations:V9D and C10R.The detection rates of 34 SNPs among the 41 SNPs were all over 91% in the Wenshang Barred chicken population.These SNPs were almost homozygous mutants without polymorphism.Eight promoters and five CpG islands were predicted in the 2 kb upstream of CDKN2A in Wenshang Barred chicken.The promoter region contained a total of 12 transcription factor binding sites,including Sp1,MEB-1,YY1,C/EBPα,AP-2α.The protein encoded by this gene was a positively charged,unstable hydrophilic protein with a molecular mass of 7.30 ku and a theoretical isoelectric point of 12.82.The predictive subcellular localization of the protein was mainly in mitochondria.The protein had no signal peptide regions and signal peptide shear sites.It had eight potential phosphorylation sites and no glycosylation sites.The protein contained a typical TRP_2 (transient receptor potential ion channelⅡ) conserved domain and its secondary and tertiary structures were mainly α-helices and random coil.In addition,it had interactions with MDM2,NPM1,USP36 proteins.The expressions of CDKN2A gene in the back skins increased from 1 to 35 days old in Wenshang Barred chicken.

The EXORDIUM(EXO) gene was identified in Arabidopsis thaliana as an brassinosteroid(BR)-responsive gene that promotes plant growth by mediating cell expansion.In order to investigate the function of EXO gene in Brassica napus and its expression pattern in different tissues at flowering stage,we used B.napus Zhongshuang 6 as the material,cloned the sequence of the coding region of the EXO gene named BnEXO,and carried out bioinformatics analysis,and used Real-time Fluorescence Quantification to determine the relative expression of BnEXO gene in B.napus in roots,stems,leaves,petals,buds,and pericarps at flowering stage.The results showed that the CDS sequence of BnEXO gene was 945 bp,BnEXO was a stable hydrophilic non-transmembrane protein,which belonged to secreted proteins and was expressed extracellularly,and the secondary structure of the protein was dominated by the random coil.The results of expression analysis in different tissues showed that the expression of BnEXO gene in different tissues was in the order of petals,pericarps,buds,stems,roots and leaves,and the highest expression was found in petals.In addition,20 B.napus EXO genes(BnaEXO),11 B.rapa EXO genes (BraEXO),and 11 B.oleracea EXO genes (BoEXO)were identified in this study based on the protein sequences of eight EXO gene family members in A.thaliana.Most proteins of gene family members were stable proteins,localized extracellularly,with amino acid lengths ranging from 271 to 411 aa,isoelectric point predictions ranging from 5.76 to 9.60,and molecular masses ranging from 28.76 to 46.21 ku.Phylogenetic analysis classified the EXO genes into five subgroups,EXOA,EXOB,EXOC,EXOD,and EXOE,with the least number of members in the EXOB subgroup.Gene structure analyses showed that most members contained only one exon and no intron,and the sequences of EXO gene family members were highly conserved.The results of cis-element analysis of the promoter region of the members in B.napus indicated that the BnaEXO genes play important roles in plant growth and development and in adversity stress.

Plant growth and production are faced with various biological and abiotic stresses,among which salt stress seriously affects the normal growth and development,quality and yield formation of plants.Plants have evolved morphological structure,physiological and biochemical reactions and genetic basis to adapt to salt stress during the long process of evolution.In terms of morphological structure,the leaves of salt-tolerant plants have waxy layer and lower stomatal density than those of salt-sensitive plants,and salt glands,microhairs,salt vesicles,and casparian strip have salt secretion or blocking functions.In terms of physiological activity regulation,on the one hand,salt-tolerant plants have high enzymatic and non-enzymatic antioxidant substances,such as SOD,CAT,phenolic substances,on the other hand,salt-tolerant plants have a high content of osmoregulatory substances,or can synthesize osmoregulatory substances under salt stress,including soluble proteins and sugars of organic substances and inorganic ions.In terms of molecular mechanism,SOS pathway is the most clearly studied ion regulation pathway,which maintains intracellular Na⁺/K⁺ balance through the synergistic action of SOS1,SOS2 and SOS3.In addition,plant hormones and carbon metabolism pathways also play an important role in the process of plant salt tolerance.This paper summarizes the research progress of salt-tolerant plants,and discusses the potential research focus and direction of salt-tolerant plants in terms of morphological structure,physiological basis,genetic molecular basis and transgenic methods in response to salt stress,which will help researchers quickly find the breakthrough point,gradually improve the mechanism system of salt-tolerant plants,and accelerate the efficient utilization of salt-tolerant plants.

In order to investigate the physiological regulation mechanism of exogenous H₂O₂ on the cotton seedlings under NaCl stress,the cotton variety Xinluzao 48 was used as the test material in an outdoor potting method.Two-factor random combinations of salt stress (NaCl,concentration gradients of 0,100,200 mmol/L) and H₂O₂ (concentration gradients of 0,0.005,0.010,0.020,0.050 mmol/L) were set,to study the change rule of fresh weight,dry weight,chlorophyll content,chlorophyll fluorescence parameters,photosynthetic gas parameters,antioxidant enzyme activities and osmotic regulation system of cotton seedlings.The results showed that exogenous H₂O₂ effectively alleviated the inhibitory effect of salt stress on the growth of cotton seedlings,increased chlorophyll content,photosynthetic gas parameters and chlorophyll fluorescence parameters of cotton seedlings,maintained the normal operation of photosynthesis of cotton seedlings and ensured the accumulation of dry matter.Meanwhile,exogenous H₂O₂ could increase the activity of antioxidant enzymes (POD,APX,CAT),accelerated the removal of ROS from cotton seedlings.Exogenous H₂O₂ reduced the electrolyte leakage rate,MDA content,the content of osmoregulation substances such as pro,free amino acid and SS,and improved the salt resistance of cotton seedlings.Among all treatment,0.020 mmol/L exogenous H₂O₂ had the best effect in alleviating the salt stress suffered by cotton seedlings.In summary,exogenous H₂O₂ improves the adaptation of cotton seedlings to salt stress by improving photosynthetic performance,keeping stable photosynthesis in cotton seedlings,and maintaining the dynamic balance between ROS production and elimination in cotton seedlings.

Salt-alkali stress has become one of the important factors restricting agricultural production in my country.Exploring the molecular mechanism of salt tolerance of crops has important theoretical and practical value for crop breeding.The purpose of this study is to clone the ZmMPI gene in corn and transform corn plants.First,qRT-PCR was used to analyze the ZmMPI expression changes in plants treated with saline-alkali solutions.Then DNAMAN software was used to perform multiple comparison analysis of the ZmMPI protein sequence.MEGA 7.0 software was used to construct a phylogenetic tree,and a series of software were used to analyze the ZmMPI protein sequence.ZmMPI performed bioinformatics analysis.Finally,molecular cloning technology was used to successfully clone the coding sequence of the ZmMPI gene,construct a plant overexpression vector,and use Agrobacterium-mediated genetic transformation method to transform the corn inbred line B104.The overexpression transgenic plants were transformed at the genome level,transcription level and protein level.Identify and analyze changes in expression levels.The results showed that the expression level of the ZmMPI gene showed an overall trend of first increasing and then decreasing after being subjected to salt-alkali stress;the ZmMPI protein sequence comparison result showed a similarity rate of 64.15%,and the phylogenetic tree showed that ZmMPI had the highest homology with Zea mays subsp.parviglumis ABA34115.1.The protein contained a protein domain Potato_inhibit,which had an α-helix,a random coil and a β-turn.It was relatively hydrophobic and had 10 predicted Potential phosphorylation sites;the identification results of the 49 transformation events obtained showed that the ZmMPI gene in 13 over-expressed transgenic lines could be expressed normally at the genome level,and the ZmMPI gene in 10 over-expressed transgenic lines could be transcribed and translated normally.Finally,10 overexpression transgenic lines capable of normal transcription and translation were obtained,laying the foundation for further exploring the molecular mechanism of ZmMPI gene in response to salt-alkali stress.

Clarifying the effect of spring limited irrigation on the root development and grain yield of winter wheat in Haihe Plain is of great significance to reduce irrigation and improve water use efficiency.This study used Shimai 22 as the test material,irrigation treatments were traditional irrigation twice at jointing and anthesis stage(W2),no irrigation(W0),and single irrigation(W1)with four irrigation-time treatments(3L,4L,5L,and 6L)based on the number of leaves unfolded in spring.The results showed that compared with W2,W0 and W1 yield decreased by 54.6% and 24.4% respectively,the irrigation yield was highest at 4L in W1,and the effect of yield composition reduction was not significant.Limited irrigation reduced the total root weight density and root length density of winter wheat.During the anthesis period, the total root weight density of W1 decreased significantly by 17.2%, while the total root weight density and root length density of W0 decreased significantly by 47.5% and 35.1%, respectively. And under W1 condition, 4L has the highest total root weight density and root length density. The vertical distribution of roots showed that reducing the frequency of irrigation increased the distribution of roots in the soil layer below 40 cm,however,with the postponement of irrigation time,the root distribution of W1 deep soil decreased and root vigor increased.Among them, during the anthesis period, 4L was significantly higher than 6L by 28.8%, 14.2%, and 36.5% in the 120—160 cm, 160—200 cm, and 200—240 cm soil layers, respectively. Correlation and path analysis showed that total root weight density and root length density at joint—anthesis period had a positive effect on yield.The direct contribution of total root length density in 3L and 4L irrigation was the largest.Generally speaking,the root mass of 4L treatment was higher at jointing-anthesis period,the deep root distribution and root activity of 40—240 cm were increased,resulting in higher spike number and kernel number,which was beneficial to alleviate the decrease of winter wheat yield at limited irrigation,it can be used as an effective way of limited irrigation for winter wheat in Haihe Plain.

In order to screen out suitable water and nitrogen combinations for watermelons in Yellow River irrigation area of Ningxia, different water and nitrogen treatments were designed to study the effects of water and nitrogen interaction on SPAD value of watermelon leaves, fruit quality,yield and nitrogen uptake and utilization. The results showed that SPAD values were higher by W1N4,W2N3,W2N4,W3N3 and W3N4 treatment,the quality was better under nitrogenous fertilizer amounts at N2 and N3.The yield was the highest under W3N4 treatment,reaching 76 565.36 kg/ha and increased by 8.34% to 37.57% compared with other treatments significantly.Followed by W3N2 and W3N3 treatment.Compared with other levels,when the irrigation water level was W1,the water use efficiency of facility watermelon irrigation was higher.Among them,the irrigation water use efficiency of W1N3 and W1N4 treatment was higher,reaching 43.91,45.32 kg/ha respectively,while it was significantly increased by 14.00% to 56.40% from other treatments.Fruit nitrogen accumulation and total nitrogen accumulation under W3N4 treatment were all the highest compared with other treatments significantly,increasing by 22.75% to 192.36% and 17.00% to 123.39% respectively compared with the other treatments.Partial factor productivity of nitrogen and nitrogen fertilizer utilization rate under W3N2 treatment were all the highest compared with other treatments significantly.Partial factor productivity of nitrogen increased by 11.00% to 343.68%separately compared with the other treatments and nitrogen fertilizer utilization rate increased by 3.34 to 10.02 percentage points compared with other treatments.The correlation analysis showed that SPAD,the center of soluble solids,Vc,yield,irrigation water use efficiency and nitrogen accumulation,were all significantly positively correlated with each other,and they were significantly negatively correlated with partial factor productivity of nitrogen and nitrogen use efficiency,the edge of soluble solids was positively correlated with nitrogen accumulation of the plants,and negatively correlated with partial factor productivity of nitrogen and nitrogen use efficiency.To sum up,the watermelon had better quality when nitrogenous fertilizer amounts were N2(80 kg/ha) and N3(160 kg/ha),the yield-increasing effect was the best under the combination of water amount for W3(2 200 m³/ha)and nitrogenous fertilizer amount for N4(240 kg/ha).The interaction between high amount of irrigation water and nitrogenous fertilizer application is beneficial to the nitrogen absorption in watermelon,and the interaction between low nitrogen application amount and high nitrogenous fertilizer amount is conducive to utilization of nitrogen fertilizer.

In order to investigate the effect of reducing nitrogen and increasing density on the yield formation characteristics of ratooning rice,hybrid rice variety Chuangliangyou 669 was used as the material to conduct two years of field experiments under conditions of three nitrogen application rates(N1:180 kg/ha;N2:153 kg/ha;N3:126 kg/ha)and two plant row spacing(M1:20.0 cm×16.7 cm;M2:16.7 cm×16.7 cm).The results showed that:reducing nitrogen reduced the leaf area index(LAI)of ratooning rice,but increasing density within a reasonable range of nitrogen reduction could increase the LAI of the main and ratooning seasons.The LAI of N1M2 and N2M2 was higher in the interaction treatments.Reducing nitrogen and increasing density both reduced the SPAD value of ratooning rice leaves,but the effect of density was not significant.Reducing nitrogen led to a decrease in dry matter weight,while increasing density could significantly increase dry matter weight.The interaction treatment of N1M2 and N2M2 had a higher dry matter weight.Reducing nitrogen reduced the yield of ratooning rice,but increasing density within a reasonable range of nitrogen reduction could increase yield.The interaction treatment of N1M2 and N2M2 had a higher yield.Reducing nitrogen significantly reduced the number of effective panicles in the main season,the total number of grains per panicle,and the regeneration rate and number of effective panicles in the ratooning season.However,increasing density had a compensatory effect on the number of panicles.Reasonable nitrogen reduction and density increase(N2M2)could coordinate the relationship among yield components and achieve higher yields.The correlation analysis showed that reasonable nitrogen reduction and density increase increased the effective number of panicles and total grains per panicle in the main season,as well as the effective number of panicles in ratooning season mainly by increasing LAI and dry matter weight of the main and ratooning season,and thereby improving the yield of ratooning rice.Overall,the nitrogen reduction and density increase treatment N2M2(nitrogen rate of 153 kg/ha,plant row spacing of 16.7 cm×16.7 cm)can save 15% nitrogen and achieve a higher yield.

Soil organic carbon and humus components are affected by soil quality,fertilization management measures and other factors.In order to clarify the regulation effect of long-term chemical fertilizer application on soil organic carbon(SOC)and soil humus components in different soil layers,a 43 a(2021)long-term fertilization experiment was conducted in Laiyang,Shandong Province.Six treatments were selected:low nitrogen fertilizer(N1),high nitrogen fertilizer(N2),high nitrogen fertilizer combined with phosphorus fertilizer(NP),high nitrogen fertilizer combined with potassium fertilizer(NK),high nitrogen combined with phosphorus and potassium fertilizer(NPK)and no fertilizer control(CK).The results showed that compared with CK,N1 could significantly increase the SOC content of 0—5 cm,with an increase of 22.84%.Single nitrogen fertilizer treatment could significantly increase the SOC content of 5—10 cm,with an increase of 20.94% and 28.60% in N1 and N2,respectively.N1 could significantly increase the SOC content of 10—20 cm,with an increase of 17.05%,while other treatments had no significant change.Compared with CK,N1 could significantly increase the content of humic acid(HA)in 10—20 cm and 20—30 cm soil layers,with an increase of 22.86% and 40.49%,respectively,while there was no significant change in 0—10 cm soil layer.NP could significantly increase the content of fulvic acid(FA)in 0—5 cm and 5—10 cm soil layers by 89.44% and 124.63%,respectively.NK could significantly increase the content of FA in 10—20 cm soil layer by 100.22%,and NPK could significantly increase the content of FA in 20—30 cm soil layer by 107.48%.N1 could significantly increase the content of humin(Hu)in 0—5 cm soil layer,with an increase of 69.34%.N2 could significantly increase the content of Hu in 5—10 cm soil layer,with an increase of 66.18%.N1 could significantly increase the content of Hu in 10—20 cm soil layer,with an increase of 79.50%,while there was no significant change in 20—30 cm soil layer.In summary,under the conditions of this experiment,long-term application of chemical fertilizers can effectively improve the fixation of soil organic carbon in non-calcareous fluvo-aquic soil and change the composition of soil humus,and the effects of different fertilization strategies are quite different.Among them,the effect of single application of nitrogen fertilizer on carbon sequestration is better.

Abstract: Tetraploid wheat is the ancestor specie of common wheat and an important food crop.Aiming to provide new resistance sources for wheat variety breeding,the resistance tetraploid wheat germplasm was explored and their resistance genes were identified.TDI-1 is a cultivated emmer wheat that has been immune to powdery mildew in the field for many years.To determine the resistance genes carried by TDI-1,and provide a theoretical basis for genetic improvement of wheat resistance,a durum wheat TDU-1 that was susceptible to powdery mildew was used to hybridize with TDI-1,and their F₁ plants,F₂ population,and F_2:3 lines were obtained.Genetic analysis of resistance was conducted on parents TDI-1,TDU-1,and their hybrid offspring that were inoculated with powdery mildew isolate E09.Then,bulked segregant analysis method combined with molecular markers was used to map the resistance gene.The results showed that TDI-1 was susceptible to E09 during the seedling stage but immune during the adult stage.F₁ plants derived from the cross of TDI-1 and TDU-1 were immune to E09 during the adult stage.The resistance of adult F₂ individuals was separated,and the ratio of resistant and susceptible plants was 3:1($χ_{3:1}^{2}$=0.11,P=0.74);the ratio of the number of homozygous resistant,separated resistant,and homozygous susceptible F_2:3 lines was 1:2:1($χ_{1:2:1}^{2}$=0.47,P=0.79),indicating that the resistance to powdery mildew in the adult stage of TDI-1 was controlled by one dominant gene,temporarily named PmTDI-1.Subsequently,a set of molecular markers was used to amplify the parents and their F₂ population,and then four markers on chromosome 2A,including Xwmc407,NRM-2AS29,NRM-2AS45 and NRM-2AS84, confirmed to be linked to PmTDI-1. PmTDI-1 was between the flanking markers NRM-2AS45 and NRM-2AS84,with genetics distances of 1.8 cM and 4.6 cM,respectively.Therefore,the adult stage powdery mildew resistance gene PmTDI-1 was preliminarily localized on chromosome 2A.This study identified a novel dominant adult-plant-resistance powdery mildew gene PmTDI-1 from tetraploid wheat TDI-1.

To investigate the regulatory effect of DEAD-box helicase 21(DDX21)on the replication of Transmissible gastroenteritis virus(TGEV).Firstly,Western Blot was utilized to analyze the effect of TGEV infection on DDX21 expression.Furthermore,we constructed eukaryotic expression plasmids of porcine DDX21 and established knockdown stable cell lines.RT-qPCR,Western Blot,Indirect immunofluorescence(IFA)and TCID₅₀ assays were used to investigate the regulatory effect of DDX21 on TGEV replication in vitro.Western Blot analysis showed that the protein levels of DDX21 were significantly up-regulated in PK-15 cells at the early stage of TGEV infection.RT-qPCR,Western Blot,IFA and TCID₅₀ experiments showed that over-expression of DDX21 significantly increased the mRNA level and protein of TGEV N in a dose-dependent manner.And the amino acids 601—784 aa of DDX21 were critical for promoting TGEV replication.Otherwise,the titer of TGEV was significantly down-regulated in DDX21 knockdown cell lines,whereas the titer of TGEV in DDX21 knockdown cell lines was reversed under the rescue experiment.This study revealed for the first time that DDX21 promotes the proliferation of TGEV and identified the key domain of DDX21 in regulating TGEV replication,which provided a theoretical a basis for future research on the function of DDX21 protein and the pathogenesis of TGEV.

The transcription factor BnHY5-2 is associated with plant stress resistance.In order to reveal the response of Brassica napus L.transcription factor BnHY5-2 to salt alkali stress in B.napus,the response of BnHY5-2 to light and salt and alkali was analyzed by transient overexpression,qRT-PCR analysis and subcellular localization.The results revealed that under light conditions,the expression level of the BnHY5-2 gene in B.napus leaves and stems was 29.22 and 3.15 fold higher,respectively,compared to dark conditions.The higher sensitivity to light in leaves suggested that they were the primary site for light signal response.Under light conditions,the expression of BnHY5-2 in leaves and stems was significantly downregulated by 53.1% and 31.0%,respectively,when B.napus was planted in Dalian coastal saline-alkali soil;after applying saline-alkali treatment under dark conditions,the expression of BnHY5-2 was downregulated by 48.2% in the stem,while the difference in expression in the leaves was not significant,indicating organ differences,indicating that the leaves had stricter requirements for light conditions.In B.napus leaves with transient overexpression of BnHY5-2,two out of six genes related to saline-alkali stress(BnNAC32 and BnGS)showed upregulation by 1.25,3.28 fold,respectively,while the other four genes(Bnamy,BnAsp,BnNHX7,BnTPS)were downregulated by 24.8%,25.4%,71.0%,and 82.0%,respectively.Meanwhile,the content of the resistance substance betaine in B.napus increased from 0.256 to 0.573 mg/g,indicating an enhancement by 1.24 fold,suggesting that the overexpression of BnHY5-2 gene could improve the saline-alkali tolerance of B.napus.Subcellular localization results showed that the transcription factor BnHY5-2 was localized in the nucleus and regulates the expression of functional genes.Therefore,BnHY5-2 is not only related to light signaling but also participates in the saline-alkali resistance of Brassica napus L.

Calmodulin-like proteins(CMLs),one of the Ca²⁺ receptors in plants,are involved in the process of plant growth and development,as well as adaptation to environmental changes.To understand the sequence characteristics of garlic CMLs and their responses to osmotic stress,AsCML15 and AsCML42 genes from garlic variety Cangshan siliuban were cloned,and their expression patterns under drought and salt stress conditions were determined.The results showed that the open reading frame of AsCML15 and AsCML42 genes were 498 and 543 bp in length,respectively,encoding 165 and 180 amino acid residues.AsCML15 and AsCML42 harbored four and three EF-hand domains,respectively.AsCML42 was closer to Arabidopsis AtCML42 and AtCML43 in evolutionary relationship,whereas AsCML15 was more closely related to Arabidopsis AtCML15 and AtCML16.Real-time Quantitative PCR technology showed that AsCML15 and AsCML42 were expressed in bulbs,leaves,and roots,and these two genes can be induced by 200 mmol/L NaCl and 15% PEG6000.The AsCML15 and AsCML42 genes may be involved in the process of garlic resisting salt and drought stress,and their biological functions can be further identified.

Glyphosate is currently the most widely used broad-spectrum herbicide.Cultivating glyphosate tolerant crops will help improve the effectiveness of chemical control on weed in farmlands,reduce the use of pesticide,and simplify preventive and control measures.To fully detect the glyphosate tolerance(GT)loci in wheat,484 germplasm resources from the Huang-huai wheat region were used to identify glyphosate toxicity.Based on the wheat 15K SNP array data,genome-wide association analysis(GWAS)was used to explore QTL related to glyphosate tolerance in wheat.The main results were as follows:the trend of changes in glyphosate tolerance of wheat varieties cultivated in different eras was slow,and the glyphosate tolerance had not significantly improved;three glyphosate tolerant wheat germplasms(including Henong 130,Jimai 782 and Taishan 23)were selected based on the phenotypic identification results of pesticide damage;seven QTL associated with the level of wheat pesticide damage were detected by GWAS,including 19 significant SNPs,distributed on wheat chromosomes 1A(0.00—30.48 Mb),1B(6.57—30.57 Mb),1D(0.00—22.98 Mb),4A(656.09—680.09 Mb),5A(508.19—532.19 Mb),6A(54.56—85.09 Mb),and 6D(12.02—36.02 Mb);the two QTL qGlyT-1A and qGlyT-6A located on wheat chromosomes 1A and 6A were the main effector sites for glyphosate tolerance in wheat,containing a total of 16 genes that may be related to glyphosate tolerance in wheat.

In order to reveal the physiological mechanism of winter rye in response to low temperature stress,the physiological changes of osmoregulatory substances and antioxidant enzyme activity in the leaves and tillering nodes of Wintergraze 70 and White BK-1 were compared during the cold hardening and rejuvenation periods.The results showed that during the cold hardening period,rye mainly improved its cold resistance by accumulating osmoregulatory substances in leaves and tillering nodes.During this period,the semi lethal temperature gradually decreased,and the semi lethal temperature of White BK-1 reached -9.93 ℃,which was 1.95 ℃ lower than Wintergraze 70 while the soil freezing.The increase of soluble sugar and soluble protein contents in the leaves and tillering nodes of White BK-1 was greater than those of Wintergraze 70,which meant these physiological pathways played an important role in improving the tolerance of White BK-1 to low temperatures in winter.Research on the rejuvenation period found that with the extension of low temperature stress time,the content of malondialdehyde first increased and then decreased.Rye resisted the low temperature of rejuvenation period by increasing the content of osmoregulatory substances,improving the activity of antioxidant enzymes in leaves and regulating the activity of antioxidant enzymes in tillering nodes.White BK-1 suffered less damage under the low temperature stress during rejuvenation period,and its malondialdehyde content was lower than Wintergraze 70 after recovery.In addition,after recovery,two varieties of rye accumulated higher contents of proline and soluble protein in the tillering nodes,which providing sufficient nutrients for growth and development after rejuvenation.

To explore the effects of different farming modes on the growth and development,photosynthesis,leaf structure and yield of maize,and to provide a theoretical basis for optimizing the cultivation measures and creating efficient planting patterns of maize in Hexi oasis irrigation area.Two tillage methods,no-tillage(NT)and conventional tillage(CT),and three planting patterns,wheat-maize intercropping(W/M),winter rapeseed-maize rotation after wheat(W-G→M),and wheat-maize rotation(W-M),were set up in the experiment,with a total of 6 treatments.The results showed that compared with CT,the plant height,stem diameter and leaf area of NT maize increased by 6.83%,4.10% and 3.97%,respectively.The dry matter quality of intercropping maize was higher than that of rotation,but the difference was not significant.The leaf pigments increased first and then decreased with the growth period,which showed that NT chlorophyll a,b and carotenoids were 11.93%,22.41% and 13.43% higher than CT,respectively,and the difference was significant.The net photosynthetic rate(Pn)and intercellular CO₂ concentration(Ci)of NT leaves were 9.17% and 3.81% higher than those of CT.The stomatal conductance(Gs)and transpiration rate(Tr)of CT treatment were 9.95% and 1.48% higher than those of NT.The leaf structure of NT maize was better,the mesophyll cells were more and arranged in order,the vascular bundles were clearly visible,the garland structure was larger,the palisade tissue and sponge tissue were rich,and the leaf thickness of NT was 2.51% thicker than that of CT,and the difference was significant.The yield of NT maize increased by 8.02% compared with CT,and the yield benefit of intercropping was greater than that of rotation(LER>1).This study found that the growth and development,leaf structure and yield of no-tillage maize were better than those of traditional tillage,and wheat intercropping maize could be promoted as the main farming mode in this area.

To improve the applicability of biochar in saline-alkali agroecosystem,the effect and microbial mechanism of modified biochar were studied.In a 2-year field experiment,common biochar(4.5 t/ha),nitrogen-rich modified biochar(7.5 t/ha)and phosphorus-rich modified biochar(15.0 t/ha)were added to investigate their impact on crop grain yield,soil physicochemical properties and soil microbial diversity.It had been observed that the addition of biochar enhanced the quality of saline-alkali soil,with nitrogen-rich modified biochar and phosphorus-rich modified biochar demonstrating more notable effects.Biochar could boost crop yield,improve soil structure and reduce soil bulk density in saline-alkali land.The effects of the three biochar types were not consistent.Among them,the application of 15.0 t/ha phosphorus-rich modified biochar showed favorable responses,with grain yield of(8.92±0.12)t/ha,representing a 110% increase compared to the control group.Biochar affected soil microbial diversity.Common biochar increased soil microbial diversity,whereas phosphorus-rich modified biochar decreased it.With the continuous addition of biochar,soil physical and chemical properties could affect the relationship between soil microorganisms and plant structure,weakening their relationships.In this study,the application of 15.0 t/ha phosphorus-rich modified biochar was recommended to improve saline-alkali agroecosystem.

To study the resistance mechanism of TaHis gene to Fusarium head blight(FHB)in wheat,the full-length coding sequence of TaHis was cloned,and the bait vector pGBKT7-TaHis was constructed,which was then used as bait for screening a yeast two-hybrid library of wheat ear induced by FHB.After obtaining the interacting proteins,yeast two-hybridization and bimolecular fluorescence complementation were further used to verify the interaction between these proteins,and RT-qPCR was used to analyze the expression pattern of TaHis interacting protein induced by FHB in resistant and susceptible cultivars respectively.The results showed that the bait vector pGBKT7-TaHis was successfully constructed,and 18 yeast monoclones were obtained on the four deficient selection medium(SD/-Leu/-Trp/-His/-Ade)after yeast two-hybrid library screening.Blast analysis showed that a total of 5 proteins were obtained,and the coding sequence of serine/arginine-rich mRNA splicing factor SR45a-like(TaSR)was identified in 6 colonies.We cloned the full-length coding region of TaSR gene from Bainong 4299 and constructed pGADT7-TaSR vector.The experiment of yeast two-hybrid showed that the yeast cells co-transformed with pGADT7-TaSR and pGBKT7-TaHis grew well and appeared blue on SD/-Leu/-Trp/-His/-Ade/ X-α-Gal/AbA,indicating that TaSR and TaHis directly interacted in yeast cells.The vectors of YC-TaHis and YN-TaSR were constructed,and the bimolecular fluorescence complementary experiments were performed.The results showed that strong fluorescence signals were generated in tobacco cells co-transferred with YC-TaHis and YN-TaSR,which further verified the interaction between TaSR and TaHis.RT-qPCR analysis of TaSR gene expression showed that TaSR expression was up-regulated in resistant cultivar Bainong 4299,while down-regulated in susceptible cultivar Bainong 607 upon FHB infection,indicating a positive correlation between TaSR expression level and FHB resistance in wheat.To sum up,the interaction between wheat TaSR and TaHis was proved,and TaSR expression level was positively correlated with FHB resistance in wheat.

In order to study the important role of zinc finger protein C₂H₂ in the stress response to soybean cyst nematode disease(SCN),lay an important foundation for further research on anti-soybean cyst nematode.The transcriptome data of SCN3 roots inoculated with Dongnong L-10(SCN-resistant)and Heinong 37(SCN-susceptible)were utilized in the present study,and the differentially expressed gene,GmC₂H₂-2like,was screened out.The localization of this gene was observed under the inverted fluorescence microscope.The gene was bioinformatically analyzed,and its overexpression vector was cloned and constructed to transform soybean hairy roots to study its resistance function against cyst nematode disease.The results showed that GmC₂H₂-2like encoded 410 amino acid residues,with a molecular weight of 45.77 ku,an isoelectric point of 8.73,and 47 phosphorylation sites,and the protein tertiary structure contained four structures,namely,α-helix(25.61%),random coil(57.32%),extended strand(13.66%),and β-turn(3.14%);the results of the subcellular localization revealed that the encoded protein was located in the nucleus;analysis of the overexpressed trans-GmC₂H₂-2like soybean hairy roots revealed that the number of cyst nematodes per unit area of the root tissue was significantly lower than that of the empty vector control,suggesting that GmC₂H₂-2like has the function of inhibiting cyst nematodes.

In order to explore the expression of uncoupling protein 3(UCP3)in different tissues of Min pig and its effect on the expression of mitochondrial-related genes and ATP synthase-related genes in Min pig preadipocytes,the back adipose tissue of 1-month-old Min pig was collected as the research material.The preadipocytes were isolated by enzymatic digestion and cultured in vitro.At the same time,axillary fat,chest fat,back fat,inguinal fat,perirenal fat and intramuscular fat were collected to construct tissue expression profiles.Real-time Quantitative PCR was used to detect the expression of MCU family genes(MCU,MICU1,MICU2),ATP synthase(ATP5B,ATP5E)and 1,4,5-trisphosphate inositol receptor type 1(ITPR1)genes and the expression of UCP3 gene in different adipose tissues by in vitro transfection of UCP3 gene overexpression vector or interference fragment.The results showed that the enzymatic digestion method could quickly obtain sufficient preadipocytes in the back adipose tissue.The cell edge had good refractive index,clear boundary,and similar morphology to fibroblasts.UCP3 gene was expressed in different adipose tissues,with the highest expression in dorsal adipose tissue and the lowest expression in abdominal fat.After overexpression of UCP3 gene,the expression levels of MCU family genes and ITPR1 genes were significantly increased,and the expression level of ATP5B gene was significantly decreased.After UCP3 gene interference,the expression levels of MCU family genes and ITPR1 genes were significantly decreased,and the expression levels of ATP synthase genes were significantly increased.The results showed that UCP3 gene was differentially expressed in different adipose tissues,which could promote the expression of MCU family genes and ITPR1 genes and inhibit the expression of ATP5B gene.

This study aimed to investigate the function of the anthocyanidin glycosyltransferase gene(UFGT)in the anthocyanin biosynthesis in common buckwheat.Guihong 1 was used as the experimental material.A UFGT gene,named FeUFGT1,was obtained by homologous alignment analysis,and cloned followed by performing on bioinformatics analysis.Overexpression vector of this gene was constructed,then transferred into Arabidopsis thaliana anthocyanin 3-O-glycosyltransferase mutant atugt78d2 to assess its function.The results revealed that the open reading frame of FeUFGT1 was 1 404 bp and encoded 467 amino acid residues with speculated molecular weight of 51.46 ku and theoretical isoelectric point of 4.97.FeUFGT1 protein contained GT-B type glycosyltransferase family domain and a PSPG-box,specific one of the plant UGT family at the C-terminus.The phylogenetic analysis showed that FeUFGT1 protein was closely related to the 3-O-glucosyltransferase of Siraitia grosvenorii UGT74AC1.The expression level of FeUFGT1 in the white-flower petals of Fengtian 1 was 3.7-fold as high as that in the red-flower petals of Guihong 1 with a significant difference.Mutant recovery experiment showed that FeUFGT1 could restore the phenotype of the Arabidopsis mutant which lacked anthocyanin accumulation.

The NAC(NAM,ATAF1/2,CUC2)gene is an important regulatory factor in the stress signal transduction network.Cloning the NAC gene in castor,studying its molecular characteristics and expression characteristics,aiming to provide data support for the potential function of the castor NAC gene.The RcNAC100-like gene of Tongbi No.5 was cloned by RT-PCR technology,and its molecular characteristics were analyzed,including bioinformatics,subcellular localization,expression patterns,and transcription activation domain analysis.The results showed that the full length of RcNAC100-like gene cDNA was 1 244 bp,including a 1 086 bp coding sequence(CDS),encoding 361 amino acids.The protein had more irregular coil and α-helix structures,and was a hydrophilic,non-secretory protein.Phylogenetic analysis showed that the RcNAC100-like protein was most closely related to NAC proteins in Manihot esculenta and Hevea brasiliensis,with highly similar motif composition and positioning.The subcellular localization of RcNAC100-like protein was consistent with the predicted results,located in the nucleus.The predicted cis-acting elements in the RcNAC100-like promoter region indicate the presence of multiple environmental response and growth-related elements.Expression pattern analysis showed that the RcNAC100-like gene had tissue-specific expression,with the highest relative expression level in the roots.Additionally,the gene could respond rapidly to adverse environments(drought,salt,cold,and ABA stress)and actively express,indicating that the RcNAC100-like gene might be a key gene in the castor's response to stress.Transcription activation assay results showed that the RcNAC100-like transcription factor has transcriptional activation activity in yeast.In summary,the RcNAC100-like gene may play an important role in the castor's resistance to adversity.

Auxin response factor(ARF)is a class of transcription factors with B3 domain,which is a direct molecule regulating auxin response and controlling gene expression.A gene,ZmARF10,which encoded ARF protein and actively participated in drought-rehydration stress response,was previously screened in maize by analyzed transcriptome data.To further research the molecular mechanism of ZmARF10 regulating drought resistance of maize,and also provide a new idea for molecular breeding of drought resistance,the gene was firstly analyzed by bioinformatics software.Secondly,Quantitative Real-time polymerase chain reaction(qRT-PCR)was used to detect the expression pattern of ZmARF10 in different tissues,under high temperature,drought,high salt,ABA and restoring treatments,and in different inbred lines.Finally,the function of ZmARF10 was analyzed using CRISPR/Cas9 technology.The results showed that ZmARF10 was located on chromosome 3 of maize,with a total length of 2 127 bp,and encoded 708 amino acids with a typical B3 domain.The upstream 2 kb region of ATG of this gene contained response elements related to methyl jasmonate,auxin,abscisic acid and low temperature.The phylogenetic tree showed that the protein encoded by the ZmARF10 gene was closely related to sorghum.qRT-PCR results showed that ZmARF10 was a constitutive expression gene,and the expression level of ZmARF10 was the highest in mature corn roots of maize.Under high temperature,drought,high salt and ABA treatments,the expression of this gene was significantly up-regulated,and the up-regulation ratio was up to 8.2 times after drought stress.After drought stress,the expression level of ZmARF10 gene was significantly higher in the drought-resistant inbred line Zheng 36 than that in the drought-sensitive inbred line B73.Investigation of Arabidopsis wild type and ARF10-deficient mutants showed that,compared with wild type,the mutant plants showed leaf wilting and even dry death,roots curled,root branch number decreased,and lateral root growth and development were hindered under drought conditions.Determination of physiological and biochemical indexes showed that the relative water content,chlorophyll content and net photosynthetic rate of the deficient mutants were significantly lower than those of the wild-type plants after drought stress,indicating that the drought resistance of Arabidopsis decreased after ARF10 gene was knocked out.

To improve seedling blast resistance of rice cytoplasmic male sterile(CMS)line Taonong1A and its maintainer Taonong1B,the Chinese local rice variety Gumei 4 was used as the donor parent harboring the broad-spectrum and durable blast resistance gene Pigm,and the Pigm-tightly-linked codominant marker T9E3 was developed and employed in marker-assisted selection(MAS)breeding practice.T9E3 showed clear and stable polymorphism between the donor and acceptor parents,amplifying a 926 bp DNA band from Gumei 4 genome,while the PCR product from either genome of Taonong1B or Taonong1A was about 2 000 bp.Greenhouse inoculation assay was performed using thirty-two Magnaporthe oryzae isolates collected from different rice areas,Gumei 4 and the improved rice lines showed much higher resistance frequency(90.63%)than that of two acceptor parents Taonong1B and Taonong1A(9.38%).Accordingly,Gumei 4 and the improved rice lines showed high-level seedling blast resistance(0 grade)but highly susceptible(8 grade)for two acceptor parents in natural nursery.Three blast resistant CMS pure lines(Taonong1A-Pigm-1—Taonong1A-Pigm-3)and the corresponding resistant maintainers(Taonong1B-Pigm-1—Taonong1B-Pigm-3)were preliminarily bred through MAS breeding,and one pair of them,Taonong1A-Pigm-2/Taonong1B-Pigm-2,was screened for potential application duo to the high-level blast resistance and elite sterility,agronomic and yield traits.Taonong1A-Pigm-2 showed complete male sterility with nearly 60% typical abortive pollens and 40% round abortive pollens,higher stigma exsertion rate(71.1%)and lower enclosed spikelet rate(33.6%),Taonong1B-Pigm-2 exhibited excellent agronomic and yield trait indexes,such as moderate sowing-heading duration(69 days),plant height(76.2 cm),main panicle length(28.1 cm),panicles per plant(15.8),spikelets per panicle(115.3)and 1000-grain weight(27.7 g)。New parent materials were created for the application of cytoplasmic male sterile(CMS)line hybrid rice breeding,and the comprehensive improvement of seedling blast resistance and characters of Taonong1A and Taonong1B were realized.

In order to study the correlation between stem traits and the formation of elasticity and density tolerance of maize,and further reveal the mechanism of lodging resistance,6 maize varieties with different lodging resistance were selected as materials,and 3 planting densities of 6.0×10⁴,7.5×10⁴ and 9.0×10⁴ plants/ha were set.The stem pull lodging angle in the field was used as the evaluation index of elasticity,and the morphological characteristics between plants and basal nodes.The internode anatomical structure,material accumulation and mechanical characteristics were analyzed.The results showed that plant height,ear height,basal internode length,thickness,epidermal thickness,crusty tissue thickness,total vascular bundle,sheath area of small vascular bundle,fresh weight per unit length,dry weight per unit length and content of each component,puncture and folding strength had significant effects on the stem pull lodging angle,among which internode diameter(r=0.521^**)and dry weight per unit length(r=0.562^**)had the greatest effects.The greater the planting density,the smaller the internode diameter,the thickness of the hard skin tissue,the total number of vascular bundles,the fresh weight per unit length,the dry weight,the content of each component,the lodging resistance,the worse the elasticity of the stem.There were significant differences in stem traits among different varieties.The elasticity related traits of Lishou 1,Chuangyu 107,Jingnongke 728 and MC278 were better than those of other varieties,and the variation amplitude was smaller with the increase of density,so the elasticity and density tolerance were stronger.Traits such as internode diameter and dry weight per unit length had significant effects on stem pull lodging angle, that was stalk elasticity,and the variation amplitude of these traits after densification determined the density tolerance of stalk.

To clear under the condition of slow release nitrogen fertilizer applying different content of maize development rule of the diurnal change of photosynthetic characteristics and growth,the light response curve was to fit,so as to farming-pastoral zone of spring maize cultivation and nitrogen utilization to provide theoretical basis.This study took Guangde 5 as the research object,and based on the long-term positioning experiment in 2018,the leaves ear position in spinning—tasseling stage of maize were mesured and analyzed under six nitrogen gradient of N 0(N0,CK), 120(N8), 180(N12), 240(N16), 300(N20),360(N24) kg/ha in 2019 and 2020. SPAD value, intercellular carbon dioxide concentration, stomatal conductivity, diurnal variation of net photosynthetic rate, transpiration rate, light response curve and dry matter accumulation rule.The results showed that with the increase of nitrogen fertilizer application, the changes of SPAD value, net photosynthetic rate, transpiration rate and stomatal conduction degree day of maize increased first and then decreased,and N16 treatment was the highest,and intercellular carbon dioxide concentration was the lowest.The changes of net photosynthetic rate,transpiration rate and stomatal conduction degree day showed unimodal curve.The light response curve fitting analysis showed that the maximum net photosynthetic rate of N16 treatment was the highest.In 2019 and 2020,N16 treatments were 37.48%,29.51%,31.85%,18.17%,37.32% and 80.04%,59.73%,50.30%,6.42%,62.51% higher than N0,N8,N12,N20 and N24 treatments,respectively.Dry matter mass accumulation in whole plant and ear were positively correlated with maximum net photosynthetic rate,apparent quantum efficiency and SPAD value.Comprehensive analysis showed that the most suitable nitrogen fertilizer application rate in western Inner Mongolia was 240 kg/ha.

In order to study the differences in yield and nitrogen utilization,transport and accumulation of different winter wheat cultivar(line)types in the Northern Huanghuai,the yield traits and nitrogen utilization efficiency of 29 and 26 winter wheat cultivars(lines)for testing in the Northern Huanghuai wheat area were investigated and analyzed in 2021—2022 and 2022—2023,respectively.Through cluster analysis,wheat was classified into four types:high-yielding,medium-high-yielding,medium-yielding and low-yielding.Nitrogen content of wheat stems,leaves,spikes,and grains at maturity were measured at anthesis and maturity,respectively,to analyze the relationship between parameters related to nitrogen utilization traits and yield.The results showed that the average yields among high-yielding,medium-high-yielding,medium-yielding and low-yielding varieties(lines)differed significantly from 2021 to 2023,with the number of spikes per hectare of high-yielding varieties significantly higher than that of other types.At the anthesis stage,the nitrogen accumulation and distribution rate of each organ were stems and sheaths>leaves>spikes;at the maturity stage,the nitrogen accumulation of each organ was grains>stems and sheaths>spikes>leaves.The post-anthesis nitrogen accumulation,pre-anthesis nitrogen transport and its contribution to the grain of different varieties of wheat were higher in high-yielding varieties(lines),and the pre-anthesis nitrogen transport and its contribution to the grain were greater than the post-anthesis nitrogen accumulation and its contribution to the grain.Nitrogen utilization efficiency,nitrogen harvest index,nitrogen accumulation at anthesis and nitrogen accumulation at maturity were significantly and positively correlated with wheat grain yield.Therefore,water and fertilizer management can be carried out through the nitrogen uptake and transfer pattern and distribution characteristics of different varieties,or selecting and breeding wheat varieties with high nitrogen utilization efficiency to achieve high-yield and high-efficiency production of wheat.

In order to explore the role of RPM1 in sorghum disease resistance,a sorghum SbRPM1 gene was obtained from sorghum smut resistant variety SX44B by homologous cloning method.The bioinformatics analysis results showed that the total length of the cDNA of SbRPM1 gene was 2 802 bp,encoding 933 amino acids,and its protein had a theoretical molecular weight of 106.1 ku and an isoelectric point of 7.11,which was a hydrophilic protein.The SbRPM1 protein had no transmembrane structure,and its subcellular localization was in the cytoplasm.Conservative domain analysis showed that SbRPM1 protein contained RX-CC-like,NB-ARC and LRR domains,and belonged to CNL proteins in the NLRs family.Phylogenetic analysis showed that SbRPM1 protein was most closely related to the RPM1 protein of Miscanthus lutarioriparius.The expression pattern of SbRPM1 gene was detected by Real-time quantitative PCR,and the results showed that the expression of SbRPM1 gene was higher in leaves and inflorescence,followed by roots,and the lowest in stem.The expression of SbRPM1 gene was significantly up-regulated at 24—72 h in disease-resistant varieties after inoculation with Sporisorium reilianum pathogen,suggesting that this gene could be induced by S.reilianum and played an important role in sorghum disease resistance.In this study, the CDS sequence of the SbRPM1 gene was cloned for the first time in sorghum, and the structure, nature and expression of the gene were characterized.

Luchuan pig myoblast determining gene 1(MyoD1)was cloned,bioinformatics analysis was performed,and its expression was analyzed in different tissues of Luchuan pigs,to explore the role of MyoD1 gene in the growth and development of skeletal muscle.Published MyoD1 gene of wild boar sequence on NCBI as the template,designed specific primer,cloned the CDS region of MyoD1,and compared the similarity with wild boar,cattle,sheep,human,mouse and rat gene sequence,constructed system tree for bioinformatics analysis by online software.Then,the relative expression of MyoD1 gene in different tissues of Luchuan pigs by RT-PCR.The CDS of MyoD1 gene in Luchuan pig was 960 bp,encoding 319 amino acids,five base mutations.The similarities of MyoD1 genes between Luchuan pig and wild boar,cattle,sheep,human,mouse and rats were 99.2%,93.0%,92.3%,90.0%,84.3% and 84.0%,respectively.Total number of MyoD1 gene atoms in Luchuan pigs was 4 680,molecular mass of the protein was 33.99 ku,molecular formula was C₁₄₅₇H₂₂₉₆N₄₄₂O₄₇₁S₁₄;the instability coefficient was 63.87,indicating a lack of stability;isoelectric point was 5.63,which belongs to the acidic protein;no transmembrane structure existed,found 35 phosphorylation sites and 1 glycosylation site;protein secondary structure was dominated by an irregular coil,which accounting for 60.69%.The MyoD1 gene in Luchuan pigs had the highest expression in the longest dorsal muscle,which was significantly higher than in other tissues,while kidney had the lowest expression.MyoD1 gene was expressed in all tissues of Luchuan pigs and mainly in the longest dorsal muscle,we speculated that MyoD1 gene might play a crucial role in the muscle growth and development of Luchuan pigs.

To clarify the effects of drought stress and cultivars on the net photosynthetic rate(Pn),the activities of photosynthetic enzymes in flag leaf in the afternoon(FLA)during grain filling stage and grain yield of winter wheat,a pond culture experiment with four water levels and two winter wheat cultivars was conducted under the condition of rainproof pond cultivation in 2019—2021.The four water treatments included severe drought(W1),moderate drought(W2),mild drought(W3),and suitable water supply(W4).The Pn,and the activities of Rubisco,RCA,PEPC,ATPase and PPDK in flag leaf from 14:00 to 16:00 during the early(EGFS)and medium(MGFS)grain filling stage,and the grain yield at maturity of strong drought resistance cultivar Jinmai 47(JM47)and weak drought resistance cultivar Yanzhan 4110(YZ4110)were investigated.Drought stress decreased the FLA Pn and activities of most photosynthetic enzymes,and the grain yield of wheat.The decrease of these indexes increased with drought stress degree,but the effects were different among varieties and years.Compared with W4,the FLA Pn of JM47 under W1,W2 and W3 decreased by 33.6%—40.6%,12.0%—30.5% and 5.0%—13.5%,as well as YZ4110 decreased by 44.0%—52.0%,22.5%—38.1% and 11.5%—20.5%,respectively.Compared with W4,the FLA Rubisco activity decreased during EGFS but increased during MGFS for JM47,while it decreased by 13.3%—25.6%,7.1%—14.0% and 11.2%—11.6% for YZ4110,respectively,under W1,W2 and W3.Compared with W4,the FLA RCA activity significantly decreased under most drought treatments during EGFS,while increased under W2 and W3 for JM47 and decreased under W1 and W2 for YZ4110 during MGFS.Compared with W4,the FLA ATPase activity of JM47decreased under W1 but increased under W3,while that of YZ4110 decreased under W1,W2 and W3 by 19.3%—48.7%,7.2%—24.2% and 0.1%—8.9%,respectively.The FLA PEPC activity under different treatments varied with growing seasons and varieties.Compared with W4,the FLA PPDK activities of JM47 and YZ4110 under W1 were decreased by 12.4%—18.8% and 16.7%—18.2%.Compared with YZ4110,in most conditions,the FLA Pn and photosynthetic enzyme activities of JM47 had no significant difference under suitable water supply(W4),but increased under drought treatments(W1,W2 and W3).The results of correlation analysis showed that yield,FLA Pn were significantly positively correlated to FLA ATPase activity during EGFS and MGFS,as well as FLA PEPC activity during EGFS.Thus,increasing the FLA ATPase and PEPC activities during the grain filling period is conducive to the increase of FLA Pn and grain yield of wheat.

The study aims to investigate the effect and mechanism of PPARA on intramuscular fat (IMF) deposition,and to search for molecular markers related to IMF deposition in sheep.The experiment was conducted using Small-tail Han sheep (STH) and a hybrid F₁ population between Suffolk sheep and Small-tail Han sheep(STH×SFK)as the research object.By measuring their meat quality,the histological structure and fat drop distribution of longissimus dorsi muscle of two sheep populations were compared by H&E staining and Oil Red O staining;qRT-PCR and WB techniques were used to detect the expression differences of PPARA mRNA and protein levels among different populations,and analyze the correlation between PPARA gene and IMF;Sanger sequencing technology was used to detect the PPARA gene SNP site to evaluate the possibility of using it as a genetic marker related to IMF deposition in sheep.The results indicated that the shear force and water loss rate of STH were significantly higher than those of STH×SFK,but pH was extremely significantly lower than that of STH×SFK,marble score of STH was lower than that of STH×SFK population;histological staining showed that STH muscle fibers were thick and tightly arranged,with IMF concentrated in the muscle fiber gaps,and muscle fibers were thin and loose in structure in STH×SFK,and IMF was evenly and widely distributed among muscle cell and muscle fiber spaces,with a higher content.The expression levels of PPARA mRNA in the STH were significantly higher than those in the STH×SFK population,and PPARA protein expressions of the STH were extremely significantly higher than those of the STH×SFK population.Correlation analysis showed that the expression level of PPARA gene was extremely significantly negatively correlated with sheep fat droplet area ratio and pH,extremely significantly positively correlated with shear force and water loss rate,and weakly correlated with marble score.Sanger sequencing results showed that base mutations T49885C,T50007C,G50013A,G50835A,G50942A and G51154C occurred in the second intron of PPARA gene in two sheep populations,but no C49885T mutation was detected in the STH population.The analysis of SNP genetic diversity showed that the homozygosity of SNP loci in the population was greater than the heterozygosity,and they were in a medium to low degree of polymorphism in the population;except for the G50835A mutation,all other SNPs were in line with the Hardy-Weinberg equilibrium,and with a stable genetic basis.It was believed that PPARA gene had an important impact on IMF deposition in sheep,and could be used as a potential molecular genetic marker for IMF trait selection in sheep.

In order to provide theoretical basis for straw fertilizing high-yield field and improving saline-alkali field in the process of continuous high yield and stable yield of spring corn in Inner Mongolia plain irrigation area.This study set up a positioning test of corn straw returning to the field for 1 to 4 years (HT1—HT4), taking the straw not returning to the field as the control (CK). Soil organic carbon content, total nitrogen content, available nitrogen content, available phosphorus content, cation exchange capacity, pH value and acid-base buffer curve were measured before spring sowing and during harvest. The results showed that, the relative change rates of soil organic carbon content, total nitrogen content, available nitrogen content, available phosphorus content and cation exchange capacity were 1.34%—3.62%, 0.20%—1.51%, -0.11%—0.78%, 0.89%—6.36% and 0.09%—0.41% during harvest period of HT1—HT4 compared with that before spring sowing. The relative change rates of soil organic carbon content, total nitrogen content, available nitrogen content, available phosphorus content and cation exchange capacity were 1.57%, -0.02%, -0.45%, -0.15% and -0.05% in CK harvest period compared with that before spring sowing. Soil pH value of HT2, HT3 and HT4 was significantly lower than that of CK. The buffer capacity of soil to alkali was HT4>HT3>HT2>HT1>CK. In summary, with the increase of straw returning years, soil carbon sequestration capacity, fertilizer retention capacity and buffering performance increase, and the ability to effectively resist drastic changes in soil pH value caused by fertilizer application and other factors was enhanced, soil quality was significantly improved by straw returning to field for fertilizer cultivation and soil improvement measures.

In order to explore the suitable phosphate fertilizer application methods in the Huang-Huai area,this research conducted field experiments to investigate the dry matter accumulation,root morphological indexes,available phosphorus content and phosphatase activity in different soil layers,maize yield and its component factors of four phosphate fertilizer application methods(conventional broadcast (P1),layered application (P2),strip application (P3) and hole application (P4)).The results showed that the ear length and kernel row number of P2 and P4 were significantly increased by 22.57%,16.81% and 15.19%,7.60% compared with P1.The grain number per spike of P2,P3 and P4 were significantly increased by 25.26%,13.86% and 17.00% compared with P1,respectively.Additionally,a decrease of spikes per area was found in P2 (15.30%) compared with P1.The yields of P2 and P4 were significantly increased by 15.20% and 10.79% compared with P1.Among root traits,the root length,root surface area,root volume and root tip number were significantly affected by the phosphorus application method.The root length under P2 and P4 treatments was significantly increased by 30.41% and 33.75% compared with P1.The root surface area of P2 treatment was significantly increased by 23.77% compared with P1.The root surface area of P4 treatment was significantly increased by 29.60% and 21.70% compared with P1 and P3 treatments,respectively.The root volume and root tip number of P2 and P4 treatments were significantly higher than those of P1 and P3.The analysis of soil available phosphorus content showed that the available phosphorus contents of P2 and P4 were significantly higher than that of P1 in the soil layer of 10—20 cm at three-leaf stage of maize.The available phosphorus contents of P2,P3 and P4 in the 0—20 cm soil layer were lower than that of P1,and the available phosphorus content of P4 in the soil layer of 20—30 cm was significantly lower than that of P1 at maturity stage.Correlation analysis showed that there was a significant positive correlation between acid phosphatase activity and soil available phosphorus content in the 10—20 cm soil layer at three-leaf stage of maize.The acid phosphatase activity and soil available phosphorus content in P2 and P4 treatments were higher in this soil layer,which was conducive to the conversion of soil phosphorus into maize absorbable state.In conclusion,compared with the traditional application method,layered phosphorus application and cave phosphorus application can improve the soil phosphorus availability in the early stage of maize,promote the growth and development of roots,and then increase the summer maize yield,so they are more suitable application method for corn phosphorus fertilizer in Huang-Huai area.

To investigate the genetic mechanism behind Annong 1687's ability to resist stripe rust,expedite the dissemination of Annong 1687 wheat variety,offer guidance for enhancing the genetics of novel wheat varieties,and mitigate the damage inflicted by wheat stripe rust on Chinese wheat production.This study employed Annong 1687(Annong 1687 is a semi-winter wheat type with resistance to the less significant physiological species CYR32.It was developed through the crossing of Annong 1106 and Xinong 822)with its sister lines,and parents.The wheat 55K SNP microarray was used to scan the entire genome of Annong 1687 as well as parents and sister lines.Concurrently,Annong 1687 and sister lines were inoculated and characterized employing the physiological race CYR32.The resistance to stripe rust grades was noted at the adult stage of the plant,and the resulting combined inoculation and field phenotypes were evaluated for stripe rust resistance of the wheat lines.The study revealed that Annong 1687 and five sister lines (lines 24,26,28,30 and 140) exhibited moderate resistance to stripe rust,while two sister lines (lines 89 and 105) displayed moderate susceptibility to this disease.Based on genomic differential data comparing Annong 1687 to its sister strains,there was evidence of enriched differential SNP loci in the 31—37 Mb segment of chromosome 2A's short arm,indicating possible presence of a gene or genes responsible for the result.To eliminate any potential interference from other stripe rust resistance genes,we verified the known stripe rust resistance genes present on the parental and chromosome 2A.Our findings indicated the possibility of a new stripe rust resistance gene in the 31—37 Mb interval of the short arm of chromosome 2A in Annong 1687.Experimental and bioinformatic analyses demonstrated that the TraesCS2A01G070700 gene exhibited disparity in the NBS structural domain of resistant and susceptible lines within this interval.Additionally,susceptible lines possessed three missense mutations within the NBS structural domain.It was hypothesised that TraesCS2A01G070700 represented a candidate gene for resistance to stripe rust in Annong 1687.

To investigate the effect of lncRNA2099 on the proliferation and differentiation of porcine preadipocytes,the qRT-PCR was used to detect the expression level of lncRNA2099 in heart,liver,spleen,lung,kidney,back muscle,back fat and proliferation and differentiation stages of porcine preadipocyte.The nucleoplasmic separation technique was used to determine the expression location of lncRNA2099 in adipocyte.The eukaryotic expression vector pcDNA3.1-lncRNA2099 was constructed and qRT-PCR was used to detect its impact on the expression of marker genes related to adipocyte proliferation and differentiation after over expression in adipocyte.The results showed that lncRNA2099 was highly expressed in kidney and back fat,but not in back muscle.The nucleoplasmic localization results showed that lncRNA2099 was expressed in both nucleus and cytoplasm;the expression levels of proliferation marker gene P21 and lipogenic differentiation marker gene LPL significantly increased after overexpression of lncRNA2099 in adipocytes.lncRNA2099 may act as a transcriptional regulator to inhibit proliferation and promote lipogenic differentiation in porcine preadipocytes.

This study aims to explore the effects of different concentrations of zearalenone (ZEN) on the growth of bovine mammary alveolar cell-T (MAC-T) and the expression of genes related to milk fat synthesis.Firstly,MAC-T cells were treated with different doses of ZEN for 36 hours,and the number of cells was counted using a hemocytometer.Then,cells were stained and analyzed for apoptosis and necrosis to determine the appropriate dose of ZEN.Next,acommercial detection kit was used to examine the impact of ZEN on reactive oxygen species (ROS) and mitochondria number in MAC-T cells.Finally,using Real-time Quantitative PCR (RT-qPCR) technology,the influence of ZEN on proliferation,apoptosis,oxidative stress,and milk fat synthesis-related gene expression in MAC-T cells was analyzed.The results showed that low doses of ZEN (0.01—1.00 μmol/L) tended to promote MAC-T growth,whereas high doses of ZEN (5.00—10.00 μmol/L) significantly reduced the number of cells,0.10 μmol/L ZEN had no obvious effect on ROS and mitochondria number in MAC-T,but 10.00 μmol/L ZEN notably elevated ROS levels and decreased mitochondria number.RT-qPCR results indicated that 0.10 μmol/L ZEN significantly promoted the expression of proliferation genes (CDK1,CCND2),antioxidant genes (DHODH,GPX4),and anti-apoptotic genes (BCL-2),but also increased the expression of apoptotic genes (CAS-3,BAX).Whereas,10.00 μmol/L ZEN significantly inhibited the expression of proliferation genes (PCNA,CDK1,CCND2),antioxidant genes (DHODH,GPX4,AIFM2),and anti-apoptotic genes (BCL-2),while significantly promoting the expression of apoptotic genes (CAS-3,BAX).Notably,0.10 μmol/L ZEN obviously promoted the expression of milk fat synthesis-related genes (PPARγ,FASN,JAK-2),but 10.00 μmol/L ZEN significantly suppressed these genes.The aforementioned results suggested different effects of ZEN concentrations on MAC-T cells:0.10 μmol/L ZEN could promote MAC-T cell growth and milk fat synthesis-related gene expression while also inducing the expression of apoptotic genes.In contrast,10.00 μmol/L ZEN induces oxidative stress in MAC-T cells,reduces mitochondria number,inhibits the expression of genes associated with proliferation,antioxidant properties,anti-apoptotic properties,and milk fat synthesis,while promoting the expression of apoptotic genes,leading to cell apoptosis.