To elucidate the mechanisms of cold response and explore superior cold-tolerance gene resources in wheat,this study employed transcriptome sequencing to uncover key regulatory networks underlying wheat cold response,and performed functional validation of the candidate gene TaGGCT18-6A.The results demonstrated that 4 ℃ cold treatments induced 10 893 and 18 784 differentially expressed genes(DEGs)in wheat seedlings after 6 h and 24 h cold treatment,respectively.KEGG analysis revealed significant enrichment of DEGs in pathways including MAPK signaling transduction and glutathione metabolism.A γ-glutamyl cyclotransferase gene TaGGCT18-6A was cloned through screening key genes in glutathione metabolism.This gene had a length of 1 772 bp,encoding 218 amino acids with conserved GGCT-like superfamily and ChaC core domain.Promoter cis-acting element analysis identified stress-responsive elements such as low-temperature-responsive(LTR)and dehydration-responsive (DRE)elements; consistently,expression pattern analysis showed sustained upregulation of TaGGCT18-6A under 4 ℃ cold treatments. Functional validation in transgenic rice revealed that overexpression lines OE#1, OE#2 and OE#3 exhibited significantly enhanced survival rate,plant height,and biomass. Overexpression lines OE#1 and OE#2 exhibited significantly enhanced plant height, while overexpression lines OE#1, OE#2 and OE#3 exhibited significantly reduced relative electrolyte leakage under -4 ℃ freezing treatments.After 4 ℃ treatment,overexpression lines accumulated higher levels of osmolytes(proline and soluble sugars),decreased malondialdehyde(MDA)content,and increased activities of superoxide dismutase(SOD),peroxidase(POD),and catalase(CAT).These findings collectively demonstrated that TaGGCT18-6A enhanced plant cold tolerance by regulating glutathione metabolism to improve antioxidant capacity.This study will provide a theoretical foundation and valuable genetic resources for molecular breeding of cold-tolerant wheat.
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To elucidate the role of WRKY transcription factor family members in the dynamic regulation of wheat growth and development,as well as in responses to abiotic stresses,this study investigated the expression patterns of TaWRKY gene under drought,high salinity,and low-temperature stress conditions.Using the common wheat cultivar Chinese Spring as the experimental material,we obtained the TaWRKY70 gene through molecular cloning.The coding sequence of TaWRKY70 was 885 bp in length,encoding a 294-amino-acid hydrophilic and unstable protein.Bioinformatics analysis revealed that the protein possessed a typical WRKYGQK conserved domain and a C2HC-type zinc finger structure,classifying it as a Group Ⅲ WRKY transcription factor.Cis-regulatory element analysis of the TaWRKY70 promoter region identified regulatory elements involved in responses to methyl jasmonate,abscisic acid,and ethylene.Co-expression gene analysis suggested that TaWRKY70 was associated with multiple stress response processes in wheat,including hormone signaling,defense against microbial pathogens,and responses to cold stress.Phylogenetic analysis indicated that TaWRKY70 shared a close evolutionary relationship with WRKY70 proteins from other Poaceae species,such as barley,maize,sorghum,and foxtail millet.Subcellular localization experiment further confirmed that TaWRKY70 was localized in the nucleus,consistent with the characteristics of a transcription factor.Expression pattern analysis showed that TaWRKY70 was expressed in wheat roots,stems,leaves,young spikes,and grains,with higher expression levels observed in roots and leaves.Under abiotic stress conditions,TaWRKY70 expression was downregulated in response to abscisic acid and low-temperature treatments but upregulated under salicylic acid,NaCl,PEG6000,and high-temperature treatments.In conclusion,the cloning of TaWRKY70 gene and analysis of its expression pattern provide a basis for the next step to analyze the molecular mechanism of TaWRKY70 involved in wheat stress resistance.
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Although high-throughput KASP markers have been developed for the wheat quality subunit 7OE,they are different from the KASP markers developed by SNP,the problem of not being able to effectively distinguish between homozygous and heterozygous remains.To clarify the issue of whether the 7OE subunit is homozygous,this study used Jinqiang 6(containing 7OE+8* subunits)and Kenong 199(containing 7+9 subunits)and hybrid offspring as materials,and used the Waxy-D1 gene of wheat as an internal reference gene.The relative copy number of the 7OE gene to the reference gene was detected by quantitative PCR using the universal dual-color fluorescence used in KASP markers to determine whether the 7OE gene exists and whether it is homozygous,and the detection results were verified by a relevant molecular marker.The results showed that the relative copy number of the parent Jinqiang 6,with the 7OE gene,was the highest,the relative copy number of the parent Kenong 199,without the 7OE gene,was 0,and the relative copy number of their hybrid F1 generation was intermediate,and the three types were easily separated.In its F2 segregating population,the relative copy numbers of the 7OE gene were also easily divided into high,medium and 0 three types.The genotypes that were detected as homozygous and heterozygous for the 7OE gene were further detected by the PCR marker of the 9 subunit(which can detect 9 subunit and the 8* subunit that are closely linked to the 7 subunit and the 7OE subunit,respectively),and the results were completely consistent.The high-throughput 7OE universal dual-color fluorescence quantitative PCR marker established in this study can accurately distinguish whether the 7OE subunit is present,and whether it is homozygous,which has a positive effect on promoting the molecular marker-assisted selection of high-quality subunit 7OE.
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OsAAP8 overexpressed transgenic lines were constructed based on the japonica rice variety Zhonghua 11.By detecting and analyzing its agronomic and quality traits,it explored the effects of OsAAP8 overexpression on rice growth and development,as well as rice quality,to provide theoretical basis for molecular design breeding by utilizing OsAAP8 gene to improve rice quality and yield.The results showed that compared with the transgenic negative plants,OsAAP8 overexpressed transgenic positive plants had significantly reduced plant height,tiller number,and single plant yield.The quality trait testing results showed that the content of glutamic acid,threonine,essential amino acids,total amino acids,protein,amylose,gel consistency,and gelatinization temperature in OsAAP8 overexpressed transgenic positive rice were significantly increased.The content of total starch and brown rice rate did not change significantly.The content of free fatty acids,taste value,polished rice rate,and whole polished rice rate were significantly decreased.The observation results of optical microscope and scanning electron microscope showed that the chalkiness rate and chalkiness degree of OsAAP8 overexpressed transgenic positive rice were significantly increased,but the chalkiness area did not change significantly,and the shape and arrangement of starch granules did not change.The grain size detection results showed no significant changes in grain length,width,and thickness of OsAAP8 overexpressed transgenic positive rice.The above results indicate that overexpression of OsAAP8 is not conducive to the growth and development of rice,but can significantly improve the nutritional quality of rice,which provides important information for the cultivation of new high-quality rice varieties.
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The TCP transcription factor family is unique to plants and plays a crucial role in key biological processes such as plant growth,metabolic activities,and stress responses.To investigate the quantity,distribution and expression patterns of TCP gene family in the Amorphophallus konjac genome,bioinformatics approaches were utilized to identify the AkTCP gene family,followed by a comprehensive analysis of the physicochemical properties,subcellular localization,collinearity,gene structure,and evolutionary relationships.Additionally,gene expression in response to biotic and abiotic stresses was examined by qRT-PCR.The findings were summarized as follows:the A.konjac genome contained 30 AkTCP family genes,with protein lengths ranging from 135 to 562 amino acids.These proteins had molecular weights between 15.08 and 57.20 ku,and isoelectric points between 4.96 and 11.49.Most of these were basic,hydrophilic,and unstable proteins,predominantly localized in the nucleus.These genes were unevenly distributed across the chromosomes.The collinear AkTCP genes were unevenly distributed across eight chromosomes,comprising four inter-chromosomal and five intrachromosomal collinearity events.The AkTCP gene structure was relatively simple,with most genes lacking introns,and all AkTCP transcription factors exhibit a highly conserved TCP domain.Phylogenetic classification separates the 30 AkTCP genes into two major classes,Class Ⅰ and Class Ⅱ,with Class Ⅱ further subdivided into the CIN and CYC/TB1 subclasses.The promoters of AkTCP genes contain cis-elements related to physiological response,light-responsive,phytohormone-responsive and stress-responsive elements.The results of qRT-PCR analysis indicated that AkTCP family members were involved in and responded positively to drought stress, salt stress, methyl jasmonate and soft rot pathogens of A. konjac.
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In order to clarify the regulatory network of oil accumulation in Brassica napus and breed oilseed rapeseed varieties with high oil content.The seed transcriptome data of four rapeseed inbred lines at 25,35,and 45 days after flowering were used to identify candidate genes affecting oil content by transcriptome analysis and correlation analysis.Consequently,a total of 1 530 genes were identified exhibiting differential expression across all three period,comprising 986 up-regulated genes and 544 down-regulated genes.GO enrichment analysis of these differentially expressed genes detected 83 lipid biosynthesis genes,79 lipid degradation genes,21 lipid transporter genes and 80 transcription factors.To further analysis of these differentially expressed transcription factors,genes including BnTT8,BnGL2,and BnNAC082 were identified.Combined with the oil content data of 50 semi-winter rapeseed in three different regions over two years,four SNPs were identified in the exons region of the BnNAC082-A03 significantly associated with oil content using genome-wide association studies.Two haplotypes were detected in the region of this gene and BnNAC082-A03_Hap1 corresponding accessions showed significantly higher oil content than that of Hap2.Additionally,it utilized these transcriptomic data to construct co-expression analysis network,and in the sub-network revealed that BnNAC082-A03 was directly connected with BnTT8-A09 and BnGL2-C06,forming a potential molecular regulatory network affecting seed lipid accumulation.
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In order to explore the relationship between MrAGL8 gene and pod dehiscence traits in Medicago ruthenica. In this study,Medicago ruthenica was used as the plant material.The MrAGL8 gene was amplified by PCR,cloned,and sequenced.Additionally,bioinformatics analysis,subcellular localization,and expression analysis in different tissues and organs were performed for this gene.The results showed that the complete coding region of MrAGL8 cDNA with a length of 711 bp was obtained through cloning using PCR amplification technology.Bioinformatics analysis results showed that MrAGL8 encoded 236 amino acids,it had MADS-box and K-box protein conserve domains.Its molecular weight was 27.39 ku,the theoretical isoelectric point was 8.69,the total number of positively charged residues was 39,the total number of negatively charged residues was 36,and the instability coefficient was 48.5.The secondary structure of the protein contained α-helix and β-sheet.It was an unstable protein and belonged to a hydrophilic alkaline protein.Subcellular localization results showed that MrAGL8 protein was located in the nucleus.The expression analysis of different tissues and organs showed that the expression level of MrAGL8 in different tissues was stem>root>pod>leaf>flower,and the expression levels of roots and stems were significantly different from those of leaves,flowers and pods.The results showed that MrAGL8 gene was related to pod dehiscence.
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To determine the relationship between the esters compounds and key enzyme activities and genes that involved in the differences in aromas of oriental melon fruits,the disparities of ester metabolites and the enzyme activities and gene expression characteristics of CXE and AAT were investigated,with the fruits of oriental melon,DX108 and DX3-5,used as experimental materials.The results showed that a total of 644 metabolites were detected in oriental melon fruits,including 114 esters.A total of 108 metabolite variations and 23 different esters were detected in mature fruits.Compared with DX3-5,the types and contents of esters in ripe fruits of DX108 increased significantly,indicating that esters were the key substances in the formation of aroma difference in ripe fruits of oriental melons.K-means clustering analysis showed that isoamyl acetate,ethyl 2-methybuyrate,β-ethyl phenylacetate,p-cresol acetate,hexyl acetate ester,methyl phenylacetate,methyl anthranilate and ethyl caproate were the key esters,which caused the difference of ripe fruit aroma between two oriental melons.In addition,the activities of AAT and CXE in fruits showed opposite trend,suggesting that the coordination of AAT and CXE affected the metabolism of esters.The expression characteristics of AAT and CXE gene family members were different with fruit development,and correlation analysis suggested that CmCXE5,CmCXE6 and CmAAT5 were involved in the metabolism of eight different esters in oriental melon ripe fruits.From those findings,the variations in synthesis and accumulation of eight key esters are important reasons that direct the difference in aroma of two kinds of oriental melon,and CmCXE5,CmCXE6 and CmAAT5 may play a major regulatory role.
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To address issues such as shallow soil layers,upward movement of the plowpan,and soil salinization in the saline-alkali soils of the West Liaohe Plain,field trials were conducted in Huatugula Town,Horqin Zouyi Middle Banner,Tongliao City,Inner Mongolia Autonomous Region,from 2020 to 2021.Two tillage methods(traditional rotary tillage and powder ridge plowing),two irrigation quotas(2 100,2 700 m3/ha),and mulching and shallow burial measures were set up,resulting in six experimental treatments:2 100 m3/ha irrigation quota+traditional rotary tillage+shallow burial(CK×NM),2 100 m3/ha irrigation quota+traditional rotary tillage+mulching(CK×DM),2 100 m3/ha irrigation quota+powder ridge plowing+shallow burial(FA×NM),2 100 m3/ha irrigation quota+powder ridge plowing+mulching(FA×DM),2 700 m3/ha irrigation quota+powder ridge plowing+shallow burial(FB×NM),2 700 m3/ha irrigation quota+powder ridge plowing+mulching(FB×DM).The effects of powder ridge plowing and mulching treatments on soil properties,structure,saline and alkaline content,and maize yield in the 0—40 cm soil layer under different irrigation quotas were analyzed. The result showed that compared to the CK×NM treatment, in the 0—40 cm soil layer, the soil bulk density decreased by 8.4%—22.9%, the total soil porosity increased by 4.9—14.8 percentage points, and the soil three-phase ratio R value decreased by 34.6%—88.2% under powder ridge plowing + mulching treatment,among them, the bulk density, total porosity, and three-phase ratio R value of the soil treated with FB×DM treatment were significantly reduced by 20.0%,-13.1 percentage points, and 88.2%, respectively;soil moisture content after sowing increased by 5.5—12.1 percentage points in the 20—40 cm soil layer, soil hardness increased by 33.4%—397.5% in the 7.5—17.5 cm soil layer,among them, the soil moisture content, hardness of the FB×DM treatment increased significantly by 12.1 percentage points, 214.3%,respectively;CO2 flux of the FB×DM treatment increased significantly by 496.4%.Compared to the CK×NM treatment, the powder ridge plowing+mulching treatment reduced the soil pH value, total alkalinity, electrical conductivity, and total salt content in the 0—40 cm soil layer, with reduction rates of 0.7%—10.9%, 2.5%—67.5%, 24.3%—68.7%, and 10.3%—81.0%, respectively. Among them, the soil pH value, total alkalinity, electrical conductivity, and total salt content of the FB×DM-treated soil were significantly reduced by 10.9%, 48.2%, 59.2%, and 80.0%, respectively.Maize germination rate, ear fresh weight, and yield were increased by 13.2—20.1 percentage points,52.5%—68.2%, and 22.4%—45.5%, respectively,among them, the germination rate, ear fresh weight, and yield of the FB×DM treatment were significantly increased by 20.1 percentage points,68.2%, and 45.5%, respectively, compared to the CK×NM treatment. Considering the comprehensive improvement effects and maize yield, it is concluded that the 2 700 m3/ha irrigation quota+powder ridge plowing+mulching(FB×DM) is a more suitable cultivation mode for saline and alkaline land in the West Liaohe Plain.
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To investigate the effects of delayed sowing on the growth,development,grain yield and quality of wheat,Yunong 907(YN907)and Yunong 922(YN922),two new high yield and high quality wheat varieties,were selected as experimental materials.Three sowing dates were established:October 22nd(S1,the conventional sowing date),October 31st(S2,delayed sowing by 9 days),and November 9th(S3,delayed sowing by 18 days).The study investigated the effects of delayed sowing on the phenological stages,grain yield and flour quality of wheat,and analyzed the relationship between temperature characteristics and wheat yield,and quality under delayed sowing conditions.The results indicated that compared with the conventional sowing date, the full growth period of YN907 and YN922 was shortened with the delay of sowing date in both years. The accumulated temperature before winter and the average daily temperature before heading decreased with the delay of sowing date, while the average daily temperature and effective accumulated temperature after heading increased. The effective accumulated temperature of the full growth period showed a downward trend. The effective accumulated temperature of the full growth period of YN907 and YN922 in S3 treatment was 243.95,222.10 ℃·d lower than that in S1 treatment in 2022—2023, and 136.30,189.40 ℃·d lower in 2023—2024, respectively. The yield and its components decreased with the delay of sowing, with a yield reduction of 6.45% to 17.26%. Compared with the conventional sowing date, the wet gluten content and protein content increased under the delayed sowing conditions.Under the background of climate warming and the scale-up of the agricultural operation,the sowing date of wheat can be adjusted to adapt to the temperature change.YN907 and YN922 had the highest yield on October 22nd.Delaying the sowing date to October 31st could maintain the higher yield level and increase the grain protein content.When the sowing date continues to be postponed,the grain protein content will be significantly increased but the yield will be significantly decreased.
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Yellow-seeded rapeseed is popular with consumers because of the appearance and good quality of rapeseed oil.However,the unstable separation of progeny traits seriously affects its large-scale application.In order to analyze the internal reasons for the unstable separation of yellow-seeded rapeseed traits,the differences in the internal physiological mechanisms between yellow and black seeds in yellow-seeded rapeseed were explored.Yellow-seeded rapeseed(CK)was used as the material to study the differences in agronomic traits,physiological and biochemical indexes,and seed coat color-related genes between yellow(Y)and black(B)seed plants in the separation offspring.The results showed that the rhizome diameter and plant height of Y were larger than those of CK and B.The plant height of B was significantly different from that of CK and Y,respectively,and the rhizome diameter of B was significantly different from that of Y.The disease index of Y was 1.97,and the disease indexes of CK and B were 2.55 and 3.33,respectively,indicating that Y was superior to CK and B in disease resistance.The content of MDA in Y leaves was the lowest at 9—10 leaf stage,and the POD activity in Y and CK flowers continued to increase at flowering stage,indicating that yellow-seeded rape had strong stress resistance.The expression levels of TT18 and TT8 genes in B and Y at 7—8 leaf stage and 9—10 leaf stage were higher than those in CK,and the expression levels of TT18 gene in B and Y at final flowering stage were significantly lower than those in CK.At 28 days after pollination,the expression of MYB47 gene in Y seed was the highest,which was 5.56 times that of CK and 5.79 times that of B,respectively.The expression of TT8 gene was the highest in Y at 21 days after pollination,which was 3.30,2.29 times that of CK and B,respectively.It showed that yellow-seeded rapeseed had obvious advantages in oil content and stress resistance.Therefore,vigorous development of yellow-seeded rapeseed can provide new ideas for improving the supply of rapeseed oil and solving the safety of edible oil in China.
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In order to explore the effects of different canopy light environments on apple mesophyll conductance and its key parameters and photosynthetic biochemical parameters,the open-center Fuji apple trees were used as experimental materials.The canopy light environment,photosynthetic biochemical parameters and leaf tissue structure of apple trees were measured after thinning,and the apple trees without thinning were used as controls(CK).Results showed that,the peak and mean value of daily maximum and averaged photosynthetically active radiation(PAR)increased by 23%,166%,and 43%,123%,respectively.Compared to the CK,product of light absorption coefficient and light energy partitioning ratio(α·β)and dark respiration rate under light(Rd)of treatment increased by 84.62%,56.00% respectively,while apparent CO2 compensation point(Ci*)value decreased by 6.67%.The average value of mesophyll conductance(gm)calculated by quantitative α·β value was 9.89% higher than that of the CK,and the maximum and average values of mesophyll conductance(gm')calculated by empirical α·β value(0.425)were 20.65%,39.38% higher than those of the CK,respectively.The gm ' of the treatment and the CK was higher than that of the gm,and the gm' of the treatment and the CK was 54.33%,21.67% higher than the average value of the gm,respectively.Vcmax derived from Pn-Ci(neglecting mesophyll conductance),Pn-Cc'(using empirical α·β)and Pn-Cc(using quantified α·β)response curves demonstrated significant differences(Vcmax-Ci and Vcmax-Cc' underestimated by 40.59%,19.85% in comparison with Vcmax-Cc in treatment,respectively),and treatment was significantly higher than CK(Vcmax-Ci,Vcmax-Cc' and Vcmax-Cc increased by 11.21%,5.85%,10.69%,respectively);Jmax and Vtpu were similar to Vcmax.The anatomical structures of leaves showed that the mean values of palisade tissue(PT)thickness and spongy tissue(ST)thickness of thinning treatment significantly increased respectively by 43.95%,43.31%,in comparison with those of the CK.The area and perimeter of PT and ST in thinning treatment were respectively 43.60% and 310.86% with 34.73% and 115.82% higher than those in CK.In summary,after thinning treatment,the light condition of apple canopy was improved,the leaf structure was changed,and the photosynthetic efficiency of the plant was improved.
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This study aimed to investigate the impacts of three exogenous amino acids on the growth and development of tomato seedlings under high-temperature stress.Tomato seedlings were selected as the study material.The normal temperature treatment with clean water at 25 ℃/18 ℃(day/night)and the high-temperature treatment with clean water at 40 ℃/30 ℃(day/night)served as control and heat groups,respectively.Under high-temperature,tomato seedlings were sprayed with 5 mmol/L γ-aminobutyric acid(GABA),0.2 mmol/L 5-aminolevulinic acid(ALA),and 5 mmol/L Proline(Pro)on the leaves once every seven days for two times.Growth parameters,antioxidant enzyme activities,and physiological parameters were determined.The results indicated that the plant height,stem diameter,and biomass accumulation of tomato seedlings was significantly reduced under high temperature.Compared with heat treatment,spraying GABA,ALA,and Pro significantly enhanced the plant height and stem thickness of tomato seedlings,promoted root development,increased biomass accumulation and relative growth rate under high-temperature.Moreover,the chlorophyll content and net photosynthetic rate of tomato seedlings declined remarkably.The chlorophyll content in the ALA treatment was higher than that in other treatments,while the net photosynthetic rate in the GABA treatment was the optimal.Compared with heat treatment,spraying GABA and ALA significantly augmented the activities of superoxide dismutase(SOD),peroxidase(POD),and catalase(CAT).High-temperature stress substantially elevated the relative electrical conductivity(REC)and malondialdehyde(MDA)content of leaves.Meanwhile,the leaves treated with Pro exhibited the lowest REC and MDA content,nearly returning to the level of control treatment.The outcomes of cluster and principal component analyses revealed that the GABA and ALA treatments mainly enhanced the photosynthetic rate and promoted plant growth by coordinately regulating the chlorophyll content and antioxidant enzyme activities of tomato seedlings,thereby strengthening the capacity to resist high-temperature stress.In contrast,Pro treatment effectively regulated the leaf's osmotic pressure by reducing leaf relative conductivity and MDA content.
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In order to study the effects of different nitrogen application rates on rice yield,nitrogen uptake and soil fertility,the effects of different nitrogen application levels on rice yield,nitrogen use efficiency and soil tillage fertility in the Yellow River Irrigation Area were studied from 2021 to 2023 using rice variety Fuyuan 4 as the test material.Compared with other treatments,the average yield of 360 kg/ha was 9.4 t/ha,which was 182.94% higher than that of no nitrogen treatment,and 40.72% and 26.34% higher than that of 210,240 kg/ha.From the perspective of yield components,the increase in yield was mainly due to the increase of grain number per spike and the number of effective spikes,and excessive nitrogen application would cause the decrease of 1000-grain weight.The average of the 3-year results showed that the nitrogen use efficiency was up to 26.93% with 240 kg/ha of nitrogen,and the partial productivity of nitrogen fertilizer decreased with the increase of nitrogen application rate.The highest average value of organic matter was 18.60 g/kg after nitrogen application of 210 kg/ha.Compared with the non-nitrogen application treatment,the soil total nitrogen content of each nitrogen application treatment increased by 7.61%—15.67%,and the soil total nitrogen content of the 360 kg/ha nitrogen application treatment decreased with the increase of the experimental year.The total phosphorus content of soil was gradually reduced in the 360 kg/ha nitrogen application treatment,and the total phosphorus content in other nitrogen application treatments was maintained at about 0.85 g/kg.The total potassium content of nitrogen fertilization treatments gradually decreased with the increase of nitrogen application rate,and the total potassium content of the 360 kg/ha nitrogen application treatment decreased by 23.74%,22.16% and 8.85%,respectively,compared with the nitrogen fertilization treatments of 120,210 and 240 kg/ha.Soil available phosphorus increased with the years of the experiment,and the variation range between treatments increased with time.With the increase of nitrogen application rate,soil available phosphorus increased first and then decreased,and the soil available phosphorus of the 240 kg/ha treatment increased significantly in 2023 compared with other treatments.The soil available potassium of each treatment did not change significantly with the test years.By considering crop yield,nitrogen use efficiency and soil fertility,nitrogen application of 240 kg/ha could balance the environment and production requirements.
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To investigate the effects of combined nitrogen and selenium fertilizer application on non-structural carbohydrates, yield, and quality of oil flax, this study utilized Longya 11 as the test material.A two-factor split-plot field experiment was conducted to examine the effects of nitrogen fertilizer(N0:0 kg/ha,N1:75 kg/ha,N2:150 kg/ha)and selenium fertilizer(Se:0 g/ha,Se1:30 g/ha,Se2:60 g/ha)on oil flax non-structural carbohydrates,grain yield and quality of oil flax under dryland conditions.The results showed that the content of soluble sugar of stems and leaves decreased gradually with the advancement of growth,while the content of starch of stems was the opposite.Under the same selenium fertilizer level,nitrogen fertilization significantly increased the soluble sugar and starch content of stems and leaves,dry matter accumulation,branch number,valid fruits,seed number grain,yield,and seed protein and oleic acid content,and the N2 level had the highest value,and the increase was greater at maturity stage.Under different nitrogen application levels,the combined application of N1 or N2 and selenium fertilizer significantly increased the soluble sugar and starch content of stems and leaves,the dry matter accumulation,branch number,valid fruits,seed number,1000-seed weight,grain yield,protein,oil content,lignans,and oleic acid content.Among the treatments,N2Se1 treatment significantly increased soluble sugar,starch content of stems and leaves and dry matter accumulation at maturity,and the valid fruits,seed number,and the 1000-grain weight were significantly higher than those of other treatments,which were 12.35%,3.89%,4.18% significantly higher than those of N2Se0 treatment,respectively.The oleic acid content of seed of N2Se1 was significantly higher than that of other treatments by 2.44%—25.64%,and the seed yield reached 1 700.50 kg/ha.The results showed that the combined application of 150 kg/ha nitrogen fertilizer and 30 g/ha selenium fertilizer could significantly increase the non-structural carbohydrate content and dry matter accumulation of oil flax stems and leaves in the test area,increase seed yield and oleic acid content,and improve quality.
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In order to explore the effect of partial replacement of chemical fertilizer by organic fertilizer on the ecological stoichiometric characteristics of soil carbon,nitrogen and phosphorus in the Loess Plateau of Eastern Qinghai,a field experiment was conducted with continuous cropping of potato(Qingshu 9).This field experiment was established in 2022 with five treatments,i.e.no fertilizer(CK),100% chemical fertilizer(T1),30% organic fertilizer+70% chemical fertilizer(T2),50% organic fertilizer+50% chemical fertilizer(T3),and 70% organic fertilizer+30% chemical fertilizer(T4).Soil contents of organic carbon,total nitrogen,total phosphorus,alkali-hydrolyzable nitrogen,available phosphorus,microbial biomass carbon,nitrogen,phosphorus and soil-microbial biomass ecological stoichiometry were determined.The results showed that compared with 2022,the contents of soil organic carbon,total nitrogen,total phosphorus,alkali-hydrolyzable nitrogen,available phosphorus and microbial biomass carbon,nitrogen and phosphorus in each fertilization treatment in 2023 showed an increasing trend,while the CK treatment showed a decreasing trend.Among different soil layers,the content of each index gradually decreased with the increase of soil depth.Among different treatments,according to the content of each index,the order was T3>T4>T2>T1>CK.In the 0—30 cm soil layer,the contents of soil organic carbon,total nitrogen,total phosphorus,alkali-hydrolyzable nitrogen,available phosphorus and microbial biomass carbon,nitrogen and phosphorus under T3 treatment were 6.88%,17.65%,17.88%,84.71%,77.01%,65.67%,80.07%,54.91% higher than those of CK,respectively.The soil C∶N and C∶P in each soil layer were the highest in the CK treatment.Among the four fertilization treatments,soil C∶N,microbial biomass C∶N and C∶P were the lowest in the T3 treatment.In addition to the 0—10 cm soil layer in 2023,the soil C∶P in each soil layer was also the lowest in T3,while the microbial biomass N∶P in the T1 treatment was the lowest among all fertilization treatments.Soil C∶N and C∶P increased with soil depth.Correlation analysis showed that organic carbon,total nitrogen,total phosphorus,available nitrogen,available phosphorus and microbial biomass carbon,nitrogen and phosphorus were significantly positively correlated with each other.In summary,the substitution of organic fertilizer for chemical fertilizer can not only change the soil nutrient and soil microbial biomass content,but also change the soil ecological stoichiometry characteristics,and the treatment with 50% organic fertilizer+50% chemical fertilizer has the best effect.
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Taking the safflower variety Yuhonghua No.1 as the research object,it adopted the planting method of two harvests of safflower and corn per year was adopted,and six fertilization treatments were set up:blank(T1),straw returning to the field(T2),straw+Bacillus subtilis bacterial fertilizer(T3(125 kg/ha),T4(250 kg/ha)),straw+Bacillus mucilaginosus bacterial fertilizer(T5(125 kg/ha),T6(250 kg/ha)).The influence of straw combined with bacterial fertilizer on the yield of safflower and soil nutrient availability was analyzed.The results showed that the continuous application of straw combined with bacterial fertilizer in 2023 and 2024 had a significant impact on the soil stoichiometric characteristics and yield.In terms of soil stoichiometric characteristics,the T5 treatment was the best for increasing total soil nitrogen(TN),the T3 treatment was the best for increasing total soil phosphorus(TP)and total soil potassium(TK),and the T4 treatment was the best for increasing soil organic matter(SOM).There were significant differences in soil N∶P,K∶P,and N∶K between two years,but the increase and decrease patterns were basically the same.The limiting factors for each treatment were all N and P elements.In terms of output,the T4 treatment was the best for increasing the filament yield,increasing by 62.53% and 52.99% respectively in two years.The T4 and T5 treatments had remarkable increasing effects on the output of seed yield.The output of T4 increased by 7.51% and 8.81% respectively in the two years,while that of T5 increased by 8.28% and 8.59%.There was an extremely significant positive correlation between soil TN,N∶P,N∶K,filament and seed yield,and there was a significant or extremely significant positive correlation between soil TN,SOM and filaments yield.Overall,among the two consecutive years of straw combined with microbial fertilizer application,the T3 treatment had the best effect in improving soil nitrogen,phosphorus,potassium and organic matter.The soils of each treatment were all limited by N and P elements.The T4 treatment had the most significant effect in increasing the comprehensive yield of filament and seed.
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To explore the expression pattern of potato gene StEXLB1a and provide a theoretical basis for the resistance to potato disease,based on the previous research of the Laboratory of Plant Resources and Molecular Biology at Northeast Agricultural University,the study using the Atlantic variety of potato as material, cloned the StEXLB1a gene, and preliminarily analyzed its bioinformatics, expression pattern, and disease resistance. The results showed that the full-length cDNA sequence of StEXLB1a gene was 768 bp,encoding 255 amino acids.The annotation indicated that it belonged to the extend protein family genes.Subcellular localization showed that the protein was localized to the cell wall.Among different tissues of potato,StEXLB1a gene expression was highest in leaves,followed by roots and stems,and lowest in flowers and tubers.The expression level of StEXLB1a gene changed significantly under various stress treatments,such as hormone(indoleacetic acid,gibberellin,abscisic acid),stress(high temperature,low temperature,salt,drought),fungal disease dry rot(Fusarium avenaceum),bacterial disease soft rot(Erwinia carotovora subsp,Ecc)and bacterial wilt(Ralstonia solanacearum,RS).The overexpressed transgenic potato plants were inoculated with the dry rot pathogen Fusarium avenaceum,and the transgenic plants were more seriously infected than the wild-type plants.The enzyme activities of active oxygen scavenging system in overexpressed plants were significantly changed,POD and SOD activities were inhibited,MDA content was higher than that of wild-type plants,and the damage degree of plants was aggravated.The results showed that transgenic potato plants with overexpression of StEXLB1a had lower resistance to dry rot than those with wild type.
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Catalase(CAT)plays a critical role in plant resistance against biotic stresses.To investigate the functions of StCAT1 and StCAT3 in potato defense against Alternaria solani infection,this study cloned the cDNA sequences of StCAT1 and StCAT3 via RT-PCR.Sequencing results demonstrated that both genes were 1 479 bp in length,encoding 493 amino acids.Phylogenetic tree construction revealed that StCAT1 shared high homology with SlCAT1 from tomato,while StCAT3 exhibited high homology with both SlCAT3(tomato)and SpCAT3 from wild tomato.Following A.solani infection for 6 days,the expression of StCAT1 and StCAT3 was significantly upregulated by approximately 3.9-fold and 8.7-fold,respectively.A co-silencing vector targeting both StCAT1 and StCAT3 was constructed,and transient co-silencing was achieved using virus-induced gene silencing(VIGS).qRT-PCR screening identified an effective co-silenced potato line(VIGS-3)with suppressed StCAT1 and StCAT3 expression.Upon A.solani inoculation,the silenced line exhibited 42% larger lesion areas compared to the control,indicating that co-silencing StCAT1/StCAT3 compromised potato resistance to early blight.DAB(3,3'-diaminobenzidine)staining further demonstrated that H2O2 accumulation in co-silenced leaves was significantly elevated,with staining intensity being 3.8-fold higher than that in the control.These results demonstrate that StCAT1 and StCAT3 regulate H2O2 homeostasis to mediate potato resistance against A.solani,providing a theoretical foundation for elucidating the molecular mechanisms of CAT genes in potato-pathogen interactions.
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To investigate the molecular mechanisms of the potato miR7997 family in response to Phytophthora infestans infection,this study analyzed the sequence characteristics,target gene prediction,expression patterns,and stress-responsive expression dynamics of Stu-miR7997 and its targets using the potato cultivar Desiree.The results revealed that the potato miR7997 family comprised three members(Stu-miR7997a/b/c)distributed across two chromosomes,with Stu-miR7997a/b sharing identical mature sequences.All precursor sequences formed canonical stem-loop secondary structures,with minimum folding free energies ranging from -37.00 to -49.50 kcal/mol,and mature sequences were located on the 5' arm.Stu-miR7997c exhibited distinct sequence length and functional element distribution compared to Stu-miR7997a/b.Promoter analysis identified light-responsive,hormone-responsive,transcription factor-binding,and stress defense-related cis-regulatory elements;target prediction identified 19 genes predominantly co-regulated by the miR7997 family.Tissue-specific expression profiling showed that Stu-miR7997a/b were highly expressed in stems,while Stu-miR7997c accumulated predominantly in roots.Upon P.infestans infection,all miR7997 members were significantly downregulated,whereas their target genes-including the transcription factor MYB92 gene,NAD(P)H-quinone oxidoreductase gene,and pectin lyase gene-were markedly upregulated.These findings suggest that the Stu-miR7997 family may indirectly modulate potato disease resistance by negatively regulating target genes.This study provides a theoretical foundation for further exploration of the miR7997 family in potato late blight resistance.
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To investigate the function of the β-glucosidase(βGlu)gene PgβGlu4 from Pyrenophora graminea,which previous studies found to be highly expressed during the infection stage,we constructed a subcellular localization vector of PCE2-EGFP-PgβGlu4 and transformed rice protoplasts,observed the fluorescence distribution and analyzed its location of existence.Simultaneously,the PgβGlu4 gene RNAi vector was constructed,and QWC protoplasts were prepared by CaCl2-PEG4000 mediated method for genetic transformation.The function of PgβGlu4 gene was studied by detecting the vegetative growth and pathogenicity of the RNAi mutants.Phylogenetic analysis of PgβGlu4 and other homologous proteins from different pathogens showed that PgβGlu4 had a closer evolutionary relationship with that from Pyrenophora tritici-repentis.The subcellular localization results showed that PgβGlu4 was mainly localized in the nucleus and cell membrane.Four PgβGlu4 gene RNAi mutants were verified by hygromycin.qRT-PCR analysis showed that the expression of PgβGlu4 gene in four RNAi mutants decreased by 66.31%,68.60%,54.37% and 69.89%,respectively,compared with the wild isolate.The colony diameter was smaller than that of the wild isolate,and their incidence rate was reduced by 56.69,52.76,47.43,and 53.30 percentage points.After infection with the mutant strain of RNAi-PgβGlu4,the relative chlorophyll content in barley leaves ranged from 30.3 to 35.0,which was significantly higher than that of the wild-type group.The effect of PgβGlu4 gene silencing on the height of barley plants before and after infection was significant compared with that of the wild-type.The results indicated that the PgβGlu4 gene was involved in the regulation of the growth,development,and pathogenicity of Pyronophora graminea.
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To explore the association between STAT3 single nucleotide polymorphism and lactation traits of sheep,414 lactating sheep were selected as test subjects,the expression of STAT3 in seven tissues including mammary gland was detected by RT-qPCR,the single nucleotide polymorphism of STAT3 was detected by sanger sequencing,STAT3 was typed by Penta-primer amplification refractory mutation system PARMS,and the effect of the nucleotide sequence variation of the gene on milk performance traits was analysed.RT-qPCR results revealed that STAT3 expression was highest in lungs,followed by liver and mammary tissues,and lowest in spleen.A total of five novel SNPs,c.22+537 C/T,c.22+658 C/T,c.318+126 C/G,c.417+360 G/A and c.417+395 T/C,were identified in the sheep STAT3,and three genotypes were detected at each SNP locus.Association analysis revealed that ewes with the genotype CC at the c.318+126 C/G locus had a higher milk fat rate than those with GG,and CC-type at the c.22+537 C/T locus,CC-type at the c.22+658 C/T locus,GG-type at the c.417+360 G/A locus,and TT-type at the c.417+395 T/C locus all had higher milk fat percentage and dry matter content.The haplotype combination H1H7 exhibited significant positive effects on milk fat percentage,dry matter content,and ash content.Additionally,the average daily milk yield of individuals carrying the haplotype combination H1H2 was increased by 0.266 kg compared to those carrying H1H7.In conclusion,the five nucleotide variation sites of STAT3 screened in this study may provide a theoretical basis for molecular selection of milk performance traits in milk sheep.
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In order to clarify the role of PRKAA1 gene in lipid deposition in goat intramuscular precursor adipocytes,the complete CDS region of the goat PRKAA1 gene was successfully amplified and cloned using RT-PCR,followed by comprehensive bioinformatics analyses. Subsequently,PRKAA1 gene overexpression vectors were constructed utilizing a double enzyme digestion approach and transfected into intramuscular precursor adipocytes.Real-time Fluorescence Quantitative PCR(RT-qPCR)was used to measure the expression level of PRKAA1 gene in the dorsal longissimus dorsi muscle in juvenile and young adults,and the expression level of PRKAA1 gene and lipid metabolism-related genes in precursor adipocytes was also detected in the process of inducing the differentiation of adipocytes. The role of PRKAA1 gene in the formation of lipid droplets was observed and investigated using oil red O staining. The results showed that the total nucleotide sequence length of goat PRKAA1 gene was 2 520 bp. Among them,the 5' untranslated region(UTR)contained 3 bp,the coding region(CDS)was up to 1 680 bp,while the 3' untranslated region(UTR)was 837 bp,and the protein encoded by this gene consisted of 559 amino acid residues. Phylogenetic analysis of the goat PRKAA1 gene revealed that sheep was the most closely related species,followed by cow,pig,dolphin,human,mouse,and rat,whereas chicken was the most distantly related species. The PRKAA1 gene was expressed at a lower level in 9 month-age than 2 month-age in the longest dorsal muscle,and expression was present throughout the induction of differentiation of precursor adipocytes and was at its highest on day 6. In experiments overexpressing the PRKAA1 gene,the content of lipid droplets in precursor adipocytes in goat muscle was significantly reduced. In addition,the expression of genes related to lipid metabolism,such as SREBP1-c,FASN,ACC,AGPAT6,PLIN1,ATGL,and ACOX1 were significantly down-regulated,and the changes were statistically significant. However,the expression levels of several genes,DGAT1,DGAT2,LPL and HSL,did not show significant changes. These results suggest that overexpression of the PRKAA1 gene might affect lipid deposition through multiple metabolic pathways,specifically,it inhibits the ab initio synthesis of fatty acids,decreases the rate of fat synthesis,and increases the rate of fat degradation. These mechanisms work together to inhibit lipid deposition in goat intramuscular precursor adipocytes.
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In order to clarify the regulatory role of marginal zone B and B1 cell specific protein(MZB1)in intramuscular adipose differentiation of goats,the study first cloned the full-lenth sequence of goat MZB1 gene and analyzed the sequence to obtain the relevant biological characteristics;then used Real-time Quantitative PCR(qPCR)to construct the expression profile of goat MZB1 in tissues and cells;finally,the goat MZB1 overexpression plasmid was transfected into the target cells,followed by differentiation induction;the effects of MZB1 overexpression on the differentiation process were evaluated through morphological analysis and qPCR.The results showed that the goat MZB1 gene sequence was 860 bp in length,including 570 bp CDS region,encoding 189 amino acids;goat MZB1 protein was not only acidic but also negatively charged,which could be stably existed in the cells.Goat MZB1 expression was detected in eight tissues,including the heart and the liver,and the highest expression was found in the spleen.The expression of MZB1 in goat intramuscular adipocytes after 72 h of oleic acid-induced differentiation was significantly higher than that before oleic acid-induced differentiation.Goat MZB1 overexpression increased the number and size of lipid droplets in goat intramuscular adipocytes;the expression levels of FASN,PPARγ,C/EBPα,SREBP1,and C/EBPβ were highly significantly upregulated.The above results suggest that MZB1 plays a positive regulatory role in goat intramuscular adipocyte differentiation,and this positive regulation may be achieved by increasing the expression of the adipose differentiation marker genes PPARγ,C/EBPα,C/EBPβ,and the fatty acid synthesis marker genes SREBP1 and FASN.
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To study the mechanism of curcumin(CUR)improving epithelial-mesenchymal transition(EMT)of bovine endometrial epithelial cells(BEND)induced by transforming growth factor-β1(TGF-β1)by regulating autophagy,different concentrations of TGF-β1(2.5,5.0 and 10.0 ng/mL)were used to induce the EMT model.Western Blot was used to detect the effects of different concentrations of curcumin(5,10 and 20 μmol/L)on the expression of the EMT-related proteins and autophagy-related proteins in BEND induced by TGF-β1.The expression levels of the EMT-related proteins(E-cadherin,N-cadherin and α-SMA)were detected by immunofluorescence.BEND was treated with autophagy inhibitor chloroquine(CQ)(10 μmol/L)to detect the expression of the EMT-related proteins and autophagy-related proteins.The expression levels of E-cadherin,Beclin-1 and LC3 proteins in BEND induced by TGF-β1 were significantly decreased,and the expression levels of N-cadherin,α-SMA and P62 proteins were significantly increased.Curcumin treatment could significantly increase the expression levels of E-cadherin,Beclin-1 and LC3 proteins,and significantly reduce the expression levels of N-cadherin,α-SMA and P62 proteins.Compared with curcumin treatment,CQ treatment significantly increased the expression levels of N-cadherin,α-SMA,P62 and LC3 proteins,and significantly decreased the expression levels of E-cadherin and Beclin-1 proteins.Curcumin can improve TGF-β1 induced BEND EMT function by promoting autophagy.
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In order to identify the molecular biological characteristics and expression patterns of Galectin-3 gene in Chinese giant salamander before and after infection,the Galectin-3 gene was amplified by RT-PCR,and its physicochemical properties and protein structure were analyzed by bioinformatics.Real-time Quantitative PCR(qRT-PCR)was used to detect the expression of Galectin-3 in different tissues of Chinese giant salamanders before and after Aeromonas hydrophila.The results showed that the length of Galectin-3 gene CDS region was 351 bp,encoding 116 amino acids,the theoretical isoelectric point(pI)was 5.87,the relative molecular weight was 13.53 ku,and it was a stable hydrophilic protein located in the cytoplasm.Structural analysis showed that Galectin-3 protein contained a GLECT domain.The phylogenetic tree showed that the Galectin-3 amino acid sequence was closely related to Rhinatrema bivittatum.qRT-PCR results showed that Galectin-3 was expressed in all tested tissues of the salamander,and the relative expression level was the highest in the skin.The relative expression levels of Galectin-3 in the liver,muscle,intestines and kidneys of Chinese giant salamander increased significantly and reached the highest value at 12 h of infection with A.hydrophila.The relative expression levels of Galectin-3 in the spleen and skin of Chinese giant salamander increased significantly and reached the highest value at 48 h of infection with A.hydrophila.In conclusion,Galectin-3 gene was actively expressed in the early stage of salamander infected by foreign bacteria,suggesting that Galectin-3 gene may be involved in the immune regulation of salamander.
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Bimonthly, Started in 1962
CN 13-1101/S
ISSN 1000-7091
CODEN: HHUOA6
Responsible Institution: Hebei Academy of Agriculture and Forestry Sciences
Sponsored by: the Academy of Agricultural Sciences and Agricultural Association of Hebei, Beijing, Tianjin, Shanxi, Henan and Inner Mongolia.
Editor-in-chief: Qiang Zhang
Edited and Published by: Editorial Department of Acta Agriculturae Boreali-Sinica
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