The hexameric Paf1 (RNA polymerase Ⅱ associated factor 1) complex is a crucial transcription regulator in eukaryotes.Paf1-regulated expression of specific genes in plants is closely related to diverse biological processes including growth,development,and stress responses.In order to get information on the responses of Paf1 to abiotic stresses in common wheat,homologous sequence searches were performed to identify all of the genes encoding each of the Paf1 subunits in the wheat genome.mCherry fusions of the wheat Paf1 subunit proteins were expressed in protoplasts and tobacco leaves for determination of protein subcellular localization by fluorescence microscopy.qRT-PCR assays were conducted to profile the expression of wheat Paf1 subunit genes in response to different abiotic stresses.The results showed that,in wheat,five of the Paf1 subunits,TaVIP3,TaVIP4,TaVIP5,TaVIP6,and TaPHP,were each encoded by one set of homeologous genes while the sixth subunit TaVIP2 was encoded by two sets.Plant VIP2 sequences had an N-terminal proline-rich region with variable length,and wheat TaVIP2 sequences had an additional glutamine-rich region.Protein subcellular localization assays revealed the nuclear localization of TaVIP2,TaVIP4,TaVIP5,and TaVIP6 proteins and the nuclear and cytoplasmic localization of TaVIP3 and TaPHP proteins.Gene expression analyses revealed similar tissue-dependent constitutive expression variations and similar stress-induced expression patterns of wheat Paf1 subunit genes.These genes coordinately responded to the stress of high temperature by expression upregulation and to the stresses of salt and drought by expression downregulation.Collectively,our results suggested the involvement of expression regulation of Paf1 subunit genes in the responses of wheat to abiotic stresses.

Preliminary transcriptomic analysis identified ZmRAV1 as a candidate gene involved in maize's response to drought stress. To further investigate its function, this study cloned the ZmRAV1 gene, conducted bioinformatics analysis of its coding sequence, and overexpressed this gene in Arabidopsis thaliana. The function of ZmRAV1 was validated by assessing the phenotypes and physiological and biochemical indices of the transgenic Arabidopsis lines under drought conditions. The results showed that the ZmRAV1 gene had a total length of 1 176 bp and encoded 389 amino acids.It had the highest proportion of irregular coils in its secondary structure and was a hydrophilic protein that did not contain signal peptides and was non transmembrane.Subcellular localization indicated that the protein was located in the nucleus.ZmRAV1 exhibited high conservation across different species.Phylogenetic analysis indicated that ZmRAV1 shares the closest evolutionary relationship with its homolog in Miscanthus sinensis, showing a high degree of homology. After drought stress treatment,the root length of Arabidopsis thaliana lines overexpressing ZmRAV1 during germination was significantly higher than that of wild-type (WT)lines.In the seedling stage,WT showed withering or even death after drought stress,while the survival rate was lower than that of overexpressing lines.Moreover,the POD and SOD activities of ZmRAV1 overexpressing lines were higher than those of WT after drought treatment,indicating that overexpression of ZmRAV1 gene could enhance Arabidopsis thaliana's resistance to drought stress.

To clarify the effects of different tillage methods on soil moisture dynamics changes and maize yield under the condition of straw return to field in the black soil area along the foothills of Daxing'anling,based on six consecutive years of tillage positioning experiments,this study analyzed the effects of seven types of tillage methods,namely,full-crushing and deep-tillage (SCD),full-crushing and shallow-tillage (SSS),full-crushing and deep-tillage (SCS),full-crushing and re-harrowing (SCR),full-crushing and rotary tillage (STR),full-crushing and no-tillage (NTS),and conventional tillage without returning straw to the field (CK)on soil moisture characteristics,water consumption,water use efficiency,and agronomic traits and yield of maize in the 0—60 cm soil layer at different growth stages of maize in each treatment.The results showed a bimodal pattern of soil quality moisture content in 2022 and 2023.0—10 cm soil layer soil quality moisture content was significantly higher than that of CK,and the NTS treatment had the highest soil quality moisture content in several periods.10—20 cm soil layer soil quality moisture content was lower than that of CK in the SSS and NTS treatments at the jointing stage,and soil quality moisture content was higher than that of CK in the 20—40 cm and 40—60 cm soil layers in 2022 and 2023 for all the treatments.In 2022 and 2023,maize plant height in all treatments except NTS treatment was significantly higher than CK at different growth stages.The SCD treatment was the tallest and the NTS treatment was the shortest at mature stage.Leaf area index (LAI) varied little among treatments at seedling stage of maize,STR treatment had the highest LAI after the jointing stage,and all treatments had significantly higher LAI than NTS treatment at the big trumpet stage.Dry matter accumulation in all treatments except for the SCS,NTS treatment was significantly higher than in CK,and dry matter accumulation was highest in the SCD treatment and lowest in the NTS treatment at mature stage.All tillage treatments increased maize yield and water use efficiency compared with CK,but the SCD treatment was significantly higher than CK.Comprehensive analysis of the indexes showed that the two tillage methods of full-crushing and deep-tillage and full-crushing and shallow-tillage were favorable to improve soil structure,maize yield and water use efficiency in the black soil area along the foothills of Daxing'anling.

In order to investigate the effects of conservation tillage techniques on soil nutrient and enzyme activity indicators,as well as ecological stoichiometry characteristics in black soil areas,this study used a 3-year positioning experiment method,with tillage (A-A) as the control,to investigate the changes in total nutrients soil organic carbon(SOC),total nitrogen (TN),total phosphorus(TP) and enzyme activities of β-D-glucosidase(βG),leucine aminopeptidase(LAP),N-1,4-acetylglucosidase(NAG),acid phosphatase(ACP)and their ecological stoichiometry characteristics in black soil under rotary tillage (B-B),conventional no tillage (C-C),and no tillage and straw mulching with original crop (0-0) treatments.The results showed that no tillage with straw mulching significantly increased soil SOC,TN,and TP content compared to tillage;except for C/N,the C/P and N/P ratios of straw mulching in no tillage with straw mulching were higher than those in tillage.The soil SOC and TN content of rotary tillage and conventional no tillage were relatively reduced compared to tillage,while the C/N,C/N,and N:P values all significantly increased in the second year.Compared with tillage,the activities of four enzymes were significantly increased under the no tillage and straw mulching treatment.In the third year,rotary tillage significantly increased ACP and βG by 22.05% and 50.00%,respectively,compared with tillage.Rotary tillage,conventional no tillage,and no tillage with straw mulching all significantly increased soil enzyme C/P.The vector angles of soil enzyme activity under the tested cultivation methods were all less than 45 degrees,indicating that soil microorganisms in the experimental area may be limited by N;the length of enzyme vector increased significantly with age,indicating an increased degree of restriction by C.The results of principal component analysis,grey relational analysis,and correlation analysis showed that no tillage with straw mulching had the most significant impact on soil nutrient content and enzyme activity.In summary,the no tillage and straw mulching with original crop technology measures have a good improvement effect on activating soil nutrients and enzyme activity,as well as maintaining soil ecological stability.

The aim was to elucidate the effects of different foliar fertilizer nitrogen application amounts on nitrogen accumulation,translocation,and utilization in maize.This experiment was conducted in 2021—2022 using a randomized block design,with maize variety Lihe 1 as the research object.No fertilization treatment (CK),conventional root fertilization treatment (CF),foliar nitrogen reduction treatment of 20% (LF1),foliar conventional nitrogen application treatment (LF2),and foliar nitrogen increase treatment of 20% (LF3) were set up to analyze the differences in nitrogen accumulation,translocation,and utilization of maize under different nitrogen fertilizer application amounts,no fertilization and conventional root fertilization.The results showed that the nitrogen accumulation in maize stems and leaves showed a trend of first increasing and then decreasing with the advancement of the growth stage,reaching the maximum value at the tasseling and silking stage.The nitrogen accumulation per plant gradually increased with the advancement of the growth stage and reached its maximum value at mature stage,the nitrogen accumulation per plant was highest in the LF2 treatment.The proportion of nitrogen distribution in leaves was highest before the silking stage;after the silking stage,the proportion of nitrogen distribution in grains gradually increased,reaching its peak at mature stage.The CK had the lowest proportion of nitrogen accumulation in grains,while the LF1 treatment had the highest proportion of nitrogen distribution in grains in 2021,and the LF3 treatment had the highest proportion of nitrogen distribution in grains in 2022.The nitrogen transport rate and the contribution rate of nitrogen transport to grains first increased and then decreased with the increase of nitrogen application amounts,the nitrogen harvest index and nitrogen utilization efficiency decreased with the increase of nitrogen application amounts;in 2021 and 2022,the nitrogen utilization efficiency of LF1 treatment,LF2 treatment,and LF3 treatment was higher than that of CF treatment,and the nitrogen utilization efficiency of LF1 treatment was the highest.The nitrogen absorption efficiency of foliar nitrogen application treatment for two years was higher than that of CF treatment.There were no significant differences in ear length,ear thickness,and ear row number among the treatments.CK had the longest bald tip length,and the row number and hundred grain weight of each nitrogen application treatment were higher than those of CK.CF treatment had the highest biological yield,while LF1 treatment had the highest grain yield and harvest index.The grain yield of each treatment was significantly higher than CK,and the harvest index decreased with increasing nitrogen application amounts.Therefore,maize can achieve better growth effects under LF1 foliar nitrogen application in the central and western regions of Inner Mongolia.

To investigate the transcriptome differences between resistant and susceptible varieties of cauliflower after inoculation with Xanthomonas campestris pv.campestris (Xcc),and to identify genes associated with cauliflower resistance to black rot disease,the susceptible variety Y1-2 and the resistant variety EC-247 of cauliflower were selected as the research subjects.Total RNA was extracted from cauliflower leaves at 0,1,3,and 5 days post-inoculation with Xcc,respectively.High-throughput parametric transcriptome sequencing was then conducted utilizing the Illumina RNA-Seq platform,followed by Real-time Quantitative PCR for validation of selected differentially expressed genes(DEGs).DEGs associated with disease resistance were screened and analyzed.The findings revealed that 6 355 genes exhibited significant differential expression between resistant and susceptible cultivars across the four time points.KEGG enrichment analysis focused on plant disease resistance pathways,identifying 47 genes involved in plant-pathogen interactions and 61 genes related to plant hormone signaling.Cluster analysis of these gene expression levels disclosed specific genes,including one CDPK,four CMLs,one PTK,one CaM,one RLK,and one SGT1 in the plant-pathogen interaction pathway,and three auxin-responsive protein genes,a TIFY gene,an indole-3-acetic acid-amido synthetase gene,two brassinazole-resistant protein genes,and a Shaggy-associated protein kinase zeta gene in the plant hormone signaling pathway.Notably,the expression of these genes was significantly higher in resistant varieties compared to susceptible ones,indicating their active response to pathogen infection at various time points.The results indicated that these differential genes might be related to disease resistance in cauliflower,which provided important genetic resources and scientific basis for molecular breeding of disease resistance in cauliflower.

The tail type of sheep is a complex trait formed by the interaction of genetic and environmental factors.circRNA is closely related to lipogenesis.To investigate the effect of circular RNA(circRNA)on the tail fat deposition of sheep,transcriptome sequencing and differential expression analysis of sheep tail fat were performed.Candidate circRNA associated with sheep tail fat were screened,and the regulatory network diagram of circRNA-miRNA-mRNA associated with sheep tail fat deposition was constructed,the selected circRNA were located,and their functions were verified.The results showed that a total of 679 differentially expressed circRNA were detected in the transcripts of adipose tissue of two different tail types of sheep,of which 422 were up-regulated and 257 down-regulated.Moreover,GO and KEGG functional enrichment analysis was performed on differentially circRNA target genes,which involved many biological development processes such as DNA metabolism,anatomical structure development,catabolic process,autophagy,carbohydrate absorption process,cell proliferation and lipid metabolism related to fat deposition.Target gene enrichment was involved in cell growth and apoptosis,cell motility,transport and catabolism,signal transduction,transcription and translation,amino acid anabolism and other functions,suggesting that these circRNA may participate in the deposition process of sheep tail fat through the above pathways.The selected differential circRNA_0018 was localized by fluorescence in situ hybridization and verified in the precursor adipocytes.The results showed that circRNA_0018 was a true and stable cytoplasmic ring molecule,and functional verification of circRNA_0018 showed that it could promote adipocyte differentiation.circRNA_0018 may be involved in the process of fat deposition and lipid metabolism in sheep.

To analyze the variability of the novel Goose astrovirus(GAstV)in Lu'an,Anhui Province and to express the VP27-VP34 fusion protein,the gout samples were collected from a farm in Lu'an.After confirming positivity via RT-PCR,the virus was isolated by passage culture in goose embryos.Then,the isolated strain was subjected to an animal regression test,whole genome amplification sequencing and genetic evolution analysis.Subsequently,the VP27-VP34 fusion protein of the isolated strain was induced and expressed,and purified recombinant protein was used to immunize 6-week-old female BALB/c mice to produce polyclonal antibodies.Serum antibody titers were assessed using agar diffusion methods,and the specificity of the polyclonal antibodies was detected by indirect immunofluorescence(IFA).The specificity of the antibody was detected by indirect immunofluorescence(IFA),and the titer of the prepared antibody was detected by the agar diffusion method.The results showed that one strain of GAstV,named AH-2021 strain,was isolated from clinical samples.The animal regression test showed obvious urate deposition on the surface of the heart and liver of goslings,and the kidney was white and swollen.Genetic evolution results revealed that AH-2021 belonged to GAstV-1,showing 98.0%—99.0% identity with other GAstV-1 strains in GenBank.The recombinant expression vector pCold-TF-VP27-VP34 was induced by IPTG to obtain the target protein,and SDS-PAGE showed that the molecular weight of the recombinant protein was about 110 ku,which mainly existed in the form of supernatant.IFA results showed that the polyclonal antibody was able to specifically recognize the GAstV,and the agar diffusion results showed that the titer of polyclonal antibody was up to 1:16.In conclusion,a strain of novel GAstV AH-2021 was isolated from gouty goslings,and animal regression tests showed that the novel Goose astrovirus was the pathogen causing gout in goslings,and a polyclonal antibody to the VP27-VP34 fusion protein was prepared.

In order to study the synergistic regulation characteristics of plant growth regulators on canopy-root traits of spring maize and to further reveal the lodging resistant mechanism of the plants,in 2021—2022,with the maize varieties Dika 159(DK159)and Xianyu 335(XY335)as experimental materials,at two planting densities of 75 000,90 000 plants/ha,plant growth regulator treatment (PGR) and water control (CK) were set up to analyze and compare the canopy structure,stem basel internode characters,root morphological characteristics and physicochemical properties of root bleeding sap under different treatments.The results showed that PGR had regulatory effects on both the maize canopy and roots.After PGR treatment,plant height,ear height and center of gravity height decreased,the mean leaf tilt angle above ear increased,light transmittance of ear layer increased by 23.59% on average,light transmittance of basel layer increased by 18.60% on average,and stem quality of basal internode was significantly improved.At the same time,the total root number,root length and root dry weight of PGR treatment increased,the root width at 10 cm below the surface increased,the root bleeding sap flow and nutrient flow increased,and the root morphological characteristics and transport capacity were obviously optimized.The flow of CTK and IAA in the root bleeding sap increased,while the flow of GA decreased.PGR effectively reduced the occurrence of stem lodging and root lodging through the synergistic regulation of canopy and root system,the field lodging rate of maize decreased from 13.43% to 6.47%,and the average yield of maize increased by 16.10%,thus achieving stable and high yield.

Light-harvesting chlorophyll a/b binding proteins are important in plant photosynthesis and abiotic stress response.To study the characteristics of GhLhcb2A1 and its expression patterns and functions in low temperature and drought response in upland cotton,full-length CDS of GhLhcb2A1 gene was cloned from the leaf cDNA of Jimian 262 by PCR.Bioinformatic analysis was conducted to learn the basic characteristics of the gene.The expression patterns and functions in low temperature and drought response were evaluated by qRT-PCR and virus-induced gene silencing.It was shown that the length of GhLhcb2A1 CDS was 798 bp,encoding 265 amino acids.GhLhcb2A1 was highly expressed in leaves and was significantly up-regulated in leaves and roots under low temperature and drought treatment.Compared with the control,its expression maximized at 3 h under low temperature and drought in leaves with 17.42 and 30.03 folds increase respectively,whereas maximized at 6 h under low temperature and 12 h under drought in roots with 11.65 and 65.04 folds respectively.Subcellular localization assay verified that GhLhcb2A1 was expressed in the chloroplasts of cells.Compared with the control plants,GhLhcb2A1 silenced plants showed a more severe phenotype of water loss and dryness under both low temperatures and drought.The accumulated malondialdehyde content in the leaves of the silenced plants was significantly higher than that of the control,while the proline content and superoxide dismutase activity were significantly lower than those of the controls,suggesting that GhLhcb2A1 silenced plant reduced the resistance to low temperature and drought.The above results implied that this gene played a positive role in regulating low temperature and drought response.

Soybean P34 protein mainly exists in soybean seeds,and its upstream promoter was likely to regulate the high expression of downstream genes in seeds.In order to further study the tissue expression pattern of soybean P34 protein gene and the regulatory activity of soybean P34 protein gene promoter,qRT-PCR was used to detect the expression of soybean P34 protein gene in soybean tissues.The 5'upstream sequence of soybean P34 protein gene(GmP34P)was cloned.The transcription initiation sites and cis-elements were analyzed by bioinformatics.The expression vector was constructed and the tobacco was transformed by Agrobacterium-mediated leaf disk method to detect GUS expression in transgenic tobacco.The results showed that the expression of P34 protein gene in soybean seeds was significantly higher than that in roots,stems,leaves and flowers.The length of GmP34P sequence obtained by cloning was 1 380 bp.Predictive analysis showed that the transcriptional start site of this sequence was base A at position 1 342,and the sequence contained a variety of cis-acting elements related to high seed expression,such as RY element,Skn-1 motif,2S seed protbanapa,etc.The plant expression vector pCAM-GmP34P containing GUS gene driven by GmP34P promoter was obtained.The positive transgenic plants were screened by hygromycin,PCR and RT-PCR.The results showed that GUS gene expression was extremely significant in transgenic tobacco seeds compared with other tissues by qRT-PCR with positive pCAM-GmP34P transgenic tobacco plants.GUS histochemical staining showed that the GmP34P promoter could regulate the high expression of downstream GUS gene in seeds.

Soil nitrogen(N)cycling enzyme activity serves as a crucial indicator for characterizing soil fertility and N transformation.To investigate the effects of long-term application of fertilizers on the soil enzyme activities correlated with N cycling in rhizosphere soil of double-cropping rice fields in southern China,our project was based on a continue 37-year fertilization localization field experiment,including four fertilization treatments:without fertilizer as a control(CK),single fertilizer(MF),rice straw residue and mineral fertilizer(RF),and 30% organic manure and 70% mineral fertilizer(OM).The activities of N cycling enzymes in the rhizosphere soil were measured,and their correlation with soil chemical properties was analyzed.The results were as follows:compared to MF and CK treatments,OM and RF treatments significantly increased the contents of total N(TN),organic carbon(SOC),ammonium N($\mathrm{NH}_{4}^{+}-\mathrm{N}$),nitrate N($\mathrm{NO}_{3}^{-}-\mathrm{N}$)and microbial biomass N(SMBN)in rhizosphere soil,and also increased rice yield.The urease(Ure)and nitrite reductase(NiR)activities of rhizosphere soil in OM and RF treatments were significantly higher than those in MF and CK treatments.The RF treatment significantly increased rhizosphere soil hydroxylamine reductase(HyR)activities compared to the other three treatments,by 21.7%,13.0%,and 8.7%,respectively.This finding shown that OM treatment significantly increased protease(Pro),nitrogenase(Nit),nitrate reductase(NR)and nitrous oxide reductase(Nos)in rhizosphere soil compared to RF,MF and CK treatments.In comparison to MF treatment,OM treatments increased Pro,Nit,NR and Nos activities in rhizosphere soil by 20.0%,26.1%,426.1% and 26.7%,respectively.Nonetheless,the activity of nitric oxide reductase(Nor)on rhizosphere soil was considerably higher in the CK treatment than in MF,RF and OM treatments.Pearson correlation analysis revealed a substantial positive correlation between soil NR,NiR,Nit,Nos,Ure,Pro and soil TN,SOC,$\mathrm{NH}_{4}^{+}-\mathrm{N}$,$\mathrm{NO}_{3}^{-}-\mathrm{N}$,SMBN as well as rice yield.Soil Nor activity was observed to have a significantly negative connection with soil TN,SOC,$\mathrm{NH}_{4}^{+}-\mathrm{N}$,$\mathrm{NO}_{3}^{-}-\mathrm{N}$,SMBN and rice yield.The findings presented above showed that soil chemical properties and yield were substantially related to rhizosphere soil N cycling enzyme activities.Redundancy analysis(RDA)showed that the first order axis could explain 93.34% of the enzyme activity in rhizosphere soil and soil $\mathrm{NO}_{3}^{-}-\mathrm{N}$,TN and SOC contents were the key factors affecting the pattern of rhizosphere soil enzyme activities.Therefore,the long-term application of organic materials such as organic manure and rice straw can enhance soil chemical and biological characteristics,stimulate soil N cycling enzyme activities,and effectively fertilize paddy soils by partially replacing chemical fertilizers.