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.
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To explore the function of key genes in photosynthesis, a functional knockout mutant (zmC4nadp-me) of ZmC4NADP-ME, the gene encoding the rate-limiting enzyme of the dark reaction of photosynthesis in maize, was obtained. Evolutionary tree analysis showed that ZmC4NADP-ME and its homologous genes exist in multiple copies in most plants, with diverse expression patterns. Phenotypic analysis revealed that the entire zmC4nadp-me plant was yellow-green, and its seedling-stage leaves dried up and died rapidly under light. Chlorophyll fluorescence analysis indicated that Y(Ⅱ) and electron transport rate ETR(Ⅱ) of photosystem Ⅱ (PSⅡ) in zmC4nadp-me decreased significantly, with little change in Y(NPQ), while the Y(NO) increased notably. Measurement of the absorption capacity (P700) of photosystem Ⅰ (PSⅠ) found that both the electron transport rate (ETR(Ⅰ)) and the actual photoelectron efficiency (Y(Ⅰ)) of zmC4nadp-me dropped substantially, and the gap widened with increasing light intensity. Under specific light intensities, Y(ND) and Y(NA) of zmC4nadp-me were greater than those of the wild type (WT). In conclusion, ZmC4NADP-ME is essential for plant growth and development. Disruption of this gene severely stresses PSⅡ, and the plant can't alleviate this stress by increasing Y(NPQ). Meanwhile, at low light intensities, the inhibition of PSⅠ may originate from the electron donor side of PSⅠ, and as the light intensity increases, the inhibition from the electron acceptor side of PSⅠ becomes a key factor.
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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.
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In order to accelerate the breeding of pink-tomato varieties,a rapid breeding strategy was used to generate pink-fruited tomato material by CRISPR/Cas9-mediated gene editing of SlMYB12.Two adjacent target sites within the first exon of SlMYB12 were selected to construct the CRISPR/Cas9 binary vector, and then the vector was introduced into red-fruited inbred line R18-10C through Agrobacterium-mediated transformation. The homozygous mutants without exogenous Cas9 were screened using specific primers and their agronomic traits and fruit nutritional quality were analyzed. Sequencing results showed that three homozygous mutants of different mutation types were obtained and all of which were frame shift mutations caused by base deletion. Compared with wild-type red fruit tomatoes, SlMYB12-edited plants grew and developed normally,and there were no significant difference in plant height,single-fruit weight,total yield per plant,fruit total soluble solid content and lycopene level,but the mature fruit showed pink and the flavonoid naringenin chalcone(NarCh)content of tomato peels was significantly reduced in MYB12-edited plants.In summary,the procedure for the generation of pink-fruited tomato plants through CRISPR/Cas9-mediated targeted mutagenesis of SlMYB12 was set up and new pink-fruited tomato germplasms with stable inheritance were obtained.
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PHR1 is a crucial factor in balancing plant disease resistance and low phosphorus stress resistance.To investigate the nature and function of the StPHR1 gene in potato and to explore the role of StPHR1 in the process of potato resistance to Alternaria solani infection,the CDS sequence of the StPHR1 gene was cloned by PCR technology using potatoes as the research material,and the structural,physicochemical properties,and phylogenetic relationships of StPHR1 were analyzed and predicted using bioinformatics software,then,the expression level of StPHR1 during the infection of potatoes by A.solani and under different hormone treatments was analyzed using qRT-PCR technology,and subcellular localization analysis of the protein was conducted using laser confocal microscopy technology.The results showed that the CDS of the StPHR1 gene was 1 353 bp,encoding 450 amino acids.The protein had a molecular formula of C2147H3399N595O711S18,a molecular weight of 49.51 ku,and a theoretical isoelectric point of 5.07,encoding a hydrophilic,unstable protein with no signal peptide and no transmembrane structure.Its secondary structure consisted mainly of random coil and α-helix.Phylogenetic tree analysis revealed that the StPHR1 protein was most closely related to Arabidopsis thaliana; conservative domain analysis revealed that the StPHR1 protein,like other PHR1s,possesses both MYB-CC and MYB conserved structural domains at its C-terminus.Relative expression analysis found that StPHR1 was significantly induced by A.solani and salicylic acid,and it was hypothesized that StPHR1 played an important role in A.solani infection of potato and in the response to salicylic acid; and the subcellular localization showed that the StPHR1 protein was localized in the nucleus.It is hypothesized that StPHR1 may regulate potato resistance to A.solani through its MYB transcription factor activity and response to salicylic acid.
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The naked seeds of zucchinii possess significant natural advantages in processing.To investigate the genetic mechanisms underlying the hull-less trait in zucchini,we utilized the hulled 17pu10 (P1) and the hull-less 17pu08 (P2) as parental strains,and constructed populations of F1 (P1×P2),F2,and BC1.The phenotypes of the zucchini seeds in the progeny population were assessed.The results showed that the number ratio of hulled seeds and hull-less seeds in the progeny population was in line with 3∶1 separation ratio.This indicated that the hull-less trait was regulated by a single gene and that the hull-less allele was recessive.Genetic mapping within this interval revealed that the gene for hull-less in zucchini was located between the markers InDel3157329 and InDel3724121,with genetic distances of 1.4,2.6 cM,respectively,while the physical distance was 0.6 Mb.The annotation and function analysis of 24 genes in the interval showed that 4 genes were directly or indirectly involved in the biosynthesis of cell wall,cellulose and lignin.Further analysis of the expression differences of 4 genes showed that only Cp4.1LG12g04350 and Cp4.1LG12g04370 had significant differences in expression levels during seed development.It was inferred that Cp4.1LG12g04350 or Cp4.1LG12g04370 was the candidate gene controlling the hull-less trait.In addition,InDel markers linked to hull-less genes were developed,which could be used as markers to identify zucchini hull-less traits,so as to accelerate the breeding of high-quality zucchini seed hull-less varieties.
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In order to reveal the expression patterns of PsbS protein in Nicotiana tabacum,the full-length NtPsbS gene sequence was cloned from cDNA of the tobacco cultivar K326,and the multiple protein sequences of NtPsbS gene and PsbS genes of seven crops including rice,tomato and soybean were compared by DNAMAN software.The phylogenetic analysis was carried out by MEGA 11 software,and the phylogenetic tree was established by neighbor-joining method.Tissue expression of NtPsbS gene in tobacco at different growth stages was detected by qRT-PCR.The plant expression vector pS1300-PsbS-GFP was constructed to find subcellular localization of the NtPsbS mature protein.Finally,the mRNA levels of NtPsbS under abiotic stresses were analyzed in K326.The results showed that the total length of tobacco NtPsbS gene was 825 bp,encoding 274 amino acids.The homology of tobacco NtPsbS protein and tomato SlPsbS protein was the highest up to 91%.The NtPsbS gene exhibited the highest mRNA levels in the leaves among the tissues,including leaves,roots,stems,seeds and other parts of K326.The mature NtPsbS protein was located in chloroplasts.For the abiotic stress,it was found that the expression level of NtPsbS was significantly increased under salt stress,cold stress and Abscisic acid(ABA)treatments.In summary,the expression level of NtPsbS was the highest in tobacco leaves at different growth stages,and it was higher when under the treatments of salt stress,cold stress and ABA,indicating that this gene may be involved in the salt and cold stress resistance and ABA metabolic pathway in tobacco,which could provide basis and reference for the functional analysis of the NtPsbS gene in future.
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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.
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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.
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To reveal the mechanism of drought resistance rice in booting stage,six single chromosome segments substitution lines (SSSLs) constructed from Oryza meridionalis and O.glumaepatula and their recipient parent Huajingxian 74 (HJX74) were used as experimental materials for potted drought treatment.Six biochemical indexes during 0,5,10 days of drought treatment and 5 days of rewatering and 11 agronomic traits after setting stage were measured,and the drought tolerance of 7 materials was comprehensivly evaluated by correlation analysis and principal component analysis.The result showed that under drought stress,there were extremely significant difference in agronomic traits among the 7 materials.There were extremely significant difference between M78-1 and HJX74 in relative panicle length,relative empty grain number and relative grain number per panicle,and there was extremely significant difference between M148 and HJX74 in relative dented grain number,there were extremely significant differences between M107 and HJX74 in relative panicle length and relative number of secondary branches; six drought-tolerant QTLs were identified at booting stage,including qRPL1-1,qRPL2-1,qRNSB2-1,qRNDG11-1,qRNEG1-1,qRGNP1-1,which were distributed on chromosomes 1,2,and 11.The superoxide dismutase (SOD) activity and peroxidase(POD) activity increased by 3.76%—18.20% and 31.88%—100.00% after 5 days of drought,while malondialdehyde (MDA)concentration decreased by 41.07%—81.65%.After 10 days of drought,SOD activity and POD activity decreased by 9.20%—48.53% and 44.74%—79.79%,while malondialdehyde (MDA)concentration was extremely significantly higher than that on the 5th day of drought.Osmoregulatory substances such as proline,soluble sugar and soluble protein continued to increase at the 5 d and 10 d drought treatment stages,and the biochemical indexes basically returned to normal level after 5 days of rewater.The comprehensive analysis revealed that the eigenvector and contribution rates of relative seed setting rate,POD activity and proline were the largest,indicating that these three indexes could better represent the drought tolerance of rice at booting stage.In conclusion,drought stress can affect agronomic traits and biochemical indexes of rice at booting stage,and rice can regulate its metabolic process in response to drought stress.
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In order to explore the difference in starch composition content and the change of rice digestion rate in rice lines. In this study, the contents of amylose (AC), total starch (TS), rapidly digestible starch (RDS), slowly digestible starch (SDS) and resistant starch (RS)in 126 indica rice strains were analyzed by enzyme digestion method in vitro. And then detected the digestion rate of the cooked rice and estimated the glycemic index (eGI) of 18 rice lines with significant differences in AC,SDS and RS. The results indicated that the starch contents of these lines differ greatly, AC was 4.29%—25.58% with an average of 10.43%, TS was 71.69%—82.45% with an average of 77.73%, RDS was 43.31%—57.47% with an average of 50.07%, SDS was 18.96%—32.56% with an average of 25.26%, and RS was 0.59%—4.87%with an average of 2.39%. There was a certain correlation between eGI values and starch content in different rice lines. The eGI values of high AC were significantly lower than low AC, but it was also found that the eGI values of Diangu 2030 with low AC and Dianpan 3429 with low SDS were also low. The eGI values of high SDS were lower than low SDS, but there were also cases where the eGI values of low SDS were also low. The eGI value of high RS was significantly lower than low RS. The rice digestion rate was fast and the sugar release was the highest of all the rice stains within 30 min after the meal, which continued to decrease after 60 min. The eGI values of the lines with high AC, SDS or RS content were generally lower than those with low starch content. The grain digestion rates were different among the tested rice lines, and it suggested that the digestion rates of rice grain could be affected by other factors except the AC, SDS and RS. These results can offer valuable references for the development of low-GI rice varieties.
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In order to investigate the effect of exogenous 24-epibrassinolide(EBR)on the physiological characteristics of Lycium barbarum seedlings under salt stress,Ningxia L. barbarum Ningqi 7 was used as the experimental material.Five treatments were set up,namely 0 mmol/L NaCl+distilled water spray(CK),150 mmol/L NaCl+distilled water spray(N0 treatment),150 mmol/L NaCl+0.005 mg/L EBR(T1 treatment),150 mmol/L NaCl+0.050 mg/L EBR(T2 treatment),and 150 mmol/L NaCl+0.500 mg/L EBR(T3 treatment).The seedling plant height,basal diameter,aboveground and underground biomass,chlorophyll content,photosynthetic parameters,antioxidant enzyme activity and the content of osmoregulatory were measured on the 7th day,14th day,and 21st day respectively.The results showed that compared with N0 treatment,T1,T2,T3 treatments significantly increased the plant height,basal diameter,aboveground and underground biomass,photosynthetic pigments in leaves,net photosynthetic rate (Pn), stomatal conductance (Gs), transpiration rate (Tr), superoxide dismutase (SOD), peroxidase (POD), catalase(CAT)activities,as well as the content of osmoregulatory in L.barbarum seedlings,intercellular CO2 concentration (Ci) and malondialdehyde (MDA) content significantly decreased,T2 treatment had the best effect.On the 21st day of salt stress,compared to the N0 treatment,the plant height,basal diameter,aboveground and underground biomass of L.barbarum seedlings increased by 23.63%,15.45%,17.70%,and 47.06%,the content of Chlorophyll a,Chlorophyll b,and Chlorophyll a+b increased by 10.68%,12.31%,and 6.57%,respectively;Pn,Tr,and Gs increased by 55.53%,27.83%,and 9.76%,respectively;the Ci value decreased by 14.42%;the activities of SOD,POD,and CAT increased by 13.23%,20.10%,and 9.31%,respectively;the MDA content decreased by 35.28%;the proline content,soluble sugar content,and soluble protein content increased by 45.17%,86.54%,and 57.00%,respectively.In summary,an appropriate concentration of exogenous 24-epibrassinolide can promote the growth of L.barbarum seedlings under salt stress,improve the photosynthetic capacity and antioxidant enzyme activity of L.barbarum seedlings,increase the content of osmoregulatory substances,alleviate the damage of salt stress to L.barbarum seedlings,and enhance the salt tolerance of L.barbarum seedlings.Among them,exogenous 0.050 mg/L EBR has the best effect.
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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.
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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.
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To provide theoretical basis and technical support for rational nitrogen fertilizer management in high-yield cultivation of hybrid wheat,this study aims to investigate the effects of nitrogen application rate on yield formation,dry matter accumulation and distribution,nitrogen absorption and utilization,as well as the nitrogen absorption ratio of wheat fertilizers in two-line hybrid wheat.From 2020 to 2021,three two-line hybrid wheat combinations and one conventional variety were used as materials.A split plot design was adopted,with nitrogen (labeled with 15N urea) treatment as the main zone and varieties as sub zones.Four nitrogen level experiments were set up at N0,N120,N180,and N240.The dry matter accumulation and distribution,plant nitrogen absorption and utilization,and grain yield of wheat organs were analyzed and measured under different treatments during the flowering and maturity stages.The results demonstrated that a highly significant effect of nitrogen application rate and combination (variety) on wheat yield and yield components.Compared with the conventional varieties Jingdong 17,the average yield of Jingmai 21 and BH9613 increased by 10.47% and 4.07% respectively,mainly due to their higher number of spikes and grains per ear.The application of nitrogen fertilizer significantly increased the number of spikes and grains per ear of wheat,but reduced the thousand grain weight.The application of nitrogen fertilizer to four varieties significantly increased the accumulation of dry matter in wheat during the flowering and maturity stages.The dry matter weight of various organs in wheat during the flowering stage was as follows:stem>leaf>spike,and during the maturity stage,it was as follows:grains>stem>spike-stalk+glume >leaf.The average values of nitrogen fertilizer agronomic utilization efficiency and nitrogen fertilizer partial productivity under different nitrogen application rates were as follows:Jingmai 21>BH9613>Jingdong 17>BH3606,which was consistent with the yield trend.The 15N atomic percentage of the four combinations (varieties),the nitrogen content from fertilizer,and the proportion of nitrogen from fertilizer all showed the following order:grains>straw,and they significantly increased with the increase of nitrogen application rate.Compared with Jingdong 17,the proportion of soil nitrogen in the three hybrid combinations (varieties) significantly increased,indicating that hybrid wheat had stronger tolerance to barrenness from the perspective of nitrogen utilization.After comprehensive consideration and analysis,the nitrogen application rate of N240 significantly increased wheat yield compared to other treatments,making it the optimal nitrogen fertilizer application rate for wheat cultivation.The comprehensive performance of the two hybrid combinations (varieties) of Jingmai 21 and BH9613 is better than the performance of the control Jingdong 17.
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To investigate the effect of applying different gradients of nitrogen reduction on leaf production capacity of Korla fragrant pear,no fertilizer treatment(CK),no nitrogen fertilizer treatment(N0),conventional fertilizer treatment(N),and three nitrogen fertilizer reduction gradients(N1,N2 and N3,which reduced nitrogen use by 10%,20%and 30%,respectively,compared with conventional fertilizer)were set up in a total of six treatments.Leaf nutrient content,net photosynthetic rate,chlorophyll fluorescence,chlorophyll content,leaf area index and yield were compared under different fertilizer application methods based on multi-year fertilizer trials.Nitrogen fertilization significantly increased leaf and branch nutrient content,leaf chlorophyll content,leaf area index,net photosynthetic rate,chlorophyll fluorescence and yield,and soluble solids and VC content in fruit.Nitrogen reduction of 10% had no significant effect on leaf and branch nutrient content,chlorophyll fluorescence,net photosynthetic rate,chlorophyll content,leaf area index and fruit soluble solids,VC,stone cells and total acid content compared to complete N application,and N reduction of 10% to 20% had no significant effect on the orchard yield and was able to maintain it at the normal range level.Net photosynthetic rate,chlorophyll fluorescence parameters,and yield were significantly and positively correlated with N,P,K,Fe,Mn,Cu,and Zn contents in leaves and branches.According to the experimental results and analysis,the recommended range of nitrogen fertilizer reduction for Korla fragrant pear of 10—12 years old is 10%—20%(240—270 kg/ha)on the basis of fully applied nitrogen(300 kg/ha)as the optimal amount of nitrogen fertilizer reduction in fragrant pear orchards.
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Cucumber powdery mildew is one of the main diseases that adversely impacts cucumber production,posing a significant challenge to its sustainable cultivation.Identifying genes related to cucumber resistance to powdery mildew can help understand the genetic principles and molecular mechanisms of cucumber resistance to powdery mildew,and provide diverse gene resources for disease resistant breeding.This study constructed F1 and F2 populations of QK×QG using cucumber resistant inbred line QK and susceptible inbred line QG as parents.Using the extreme trait mixed pool resequencing (BSA-seq) method,the genomic regions harboring the resistance genes of cucumber powdery mildew were preliminarily located.By integrating transcriptome data with gene annotation information,the association interval of the disease phenotype was narrowed,sequence variations were identified,and key genes were screened.The results showed that the resistance of powdery mildew may be controlled by recessive genes,and the population of F2 showed a continuous normal distribution from resistant to susceptible.The BSA-seq analysis,combining the SNP-Index method and QTG(quantitative trait genomics)-seq method analysis highlighted the 19—21 Mb region of chromosome 5,where there were 77 annotated genes with SNP differences between samples,including 33 non-synonymous mutations.The transcriptome sequencing (RNA-seq) results showed that there were 309 upregulated genes and 697 downregulated genes in the susceptible material.The expression levels of 13 genes within the candidate segment of chromosome 5 showed significant differences after infection.Through a comprehensive analysis of differentially expressed genes and BSA,the candidate genes in this segment were narrowed to 3,and only SNP mutations were detected in the LOC101207011 gene.The candidate gene LOC101207011 was characterized by a mutation resulting in an amino acid change from Valine at position 656 to Leucine.This gene emerges as the primary focus of our investigation due to its potential role in conferring resistance to powdery mildew.
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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.
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Cabbage fusarium wilt is a serious soil-borne fungal disease caused by Fusarium oxysporum f.sp.conglutinans (FOC),which affects the yield and quality of cabbage.In order to clarify the biological function of the transcription factor SNT2 in this pathogen,the SNT2 gene knockout mutant ΔSNT2 in F.oxysporum was successfully obtained by homologous recombination and protoplast transformation,and its phenotype and pathogenicity were analyzed.The results showed that SNT2 in F.oxysporum encoded 1 529 amino acids,had a SANT domain that binds to DNA,and the protein belonged to a hydrophilic protein.Compared with the wild type strain,the mycelial growth rate of the ΔSNT2 mutant decreased and the septation increased significantly,and the conidial production decreased significantly.Based on the results of exogenous stress,ΔSNT2 was insensitive to osmotic stress of 1 mol/L sorbitol,but its tolerance to oxygen stress,salt stress and cell wall stress of 0.1% H2O2,2 mol/L NaCl and 0.05% Congo red was reduced.At the same time,the pathogenicity test showed that the disease index of ΔSNT2 mutant was significantly lower than that of wild type,and the deletion of SNT2 resulted in a significant decrease in the pathogenicity of F.oxysporum.In conclusion,the transcription factor SNT2 plays an important role in maintaining the integrity of the cell wall during the interaction between the pathogen and the host,and participates in the regulation of the growth and development of F.oxysporum and the expression of pathogenicity.
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This study evaluates the efficacy of bio-organic fertilizer in managing Verticillium wilt in sunflowers,examining its effects on plant growth and disease resistance,and exploring its potential as a sustainable disease control strategy in sustainable agriculture.Pot experiment was conducted to investigate the effects of bio-organic fertilizer at different concentrations and under various treatment conditions on the sunflower Verticillium wilt pathogen and its impact on sunflower growth and disease resistance.The results indicated that the treatment significantly reduced the disease index of Verticillium wilt by 14.57% compared to the control group,with a relative control efficacy of 28.54% under indoor conditions.The application of bio-organic fertilizer significantly promoted sunflower growth and development.Specifically,when 1 g of bio-organic fertilizer was mixed with every 50 g of field soil,the sunflower emergence rate increased by 8.67 percentage point,and key physiological parameters,including seedling height,stem diameter,and fresh weight,were significantly enhanced.Further investigation revealed that bio-organic fertilizer fermentation filtrates at various concentrations effectively inhibited the growth of Verticillium dahliae colonies and spore germination,with inhibitory effects diminishing as dilution increased.Additionally,volatile compounds from the fertilizer significantly suppressed the formation of microsclerotia.In terms of disease resistance mechanisms,the bio-organic fertilizer fermentation filtrates enhanced plant resistance by inducing induced resistance.Physiological analysis showed that the filtrates triggered reactive oxygen species (H2O2)bursts,elevated the activities of superoxide dismutase (SOD) and peroxidase (POD),reduced malondialdehyde (MDA) accumulation,and increased the activity of phenylalanine ammonia-lyase (PAL),thereby significantly enhancing disease resistance in sunflowers.This study highlights the control potential of bio-organic fertilizer for managing sunflower Verticillium wilt and promoting plant health,providing theoretical and experimental support for optimizing fertilizer formulations and developing effective disease control strategies to enhance agricultural production.
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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.
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Elongation of very long chain fatty acids protein 3(ELOVL3),mainly involved in the synthesis of very long chain fatty acids(VLCFA),plays an important role in fatty acid metabolism.The objectives of this study were to clone the protein-coding sequence of ELOVL3 gene from Jiulong yak,to construct the eukaryotic expression vector of ELOVL3 gene,and to predict and analyze its biological functions.The CDS region of the ELOVL3 gene was amplified by PCR and inserted into the pcDNA3.1 vector by homologous recombination to construct a recombinant plasmid.The recombinant plasmid was validated by restriction digest and PCR,and the biological functions of its protein coding sequence were analyzed by online prediction software in combination with sequencing results.In addition,qPCR and Western Blot were performed for the expression of ELOVL3 gene at mRNA level and protein level.The results showed that the protein-coding sequence of Jiulong yak ELOVL3 gene was 813 bp in length,encoding a total of 270 amino acids.Enzymatic cleavage,PCR and sequencing identification confirmed the successful construction of the pcDNA3.1-ELOVL3 eukaryotic expression vector.Bioinformatics analysis showed that the protein encoded by the ELOVL3 gene was a hydrophobic protein,with 6 transmembrane structural domains and 27 phosphorylation sites.The phylogenetic tree showed that the closest relation to the Jiulong yak was the common cow,and the furthest was the original chicken.In addition,the ELOVL3 gene was most highly expressed in the lung tissue of the Jiulong yak.
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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.
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Bone morphogenetic protein 6 gene(bmp6) plays an important role in regulating the formation and development of intermuscular bones in fish.In order to explore the sequence characteristics and expression characteristics of bmp6 gene in Carassius auratus var.Qihe,the coding sequence(CDS) of bmp6 gene was cloned and analyzed by bioinformatics.Through Quantitative Real-time PCR (qRT-PCR) detection,the expression differences of bmp6 gene in different tissues and developmental stages were identified.The results showed that the bmp6 gene of C.auratus var.Qihe was composed of two homologous genes,each with three alleles,namely bmp6a-1,bmp6a-2,bmp6a-3 and bmp6b-1,bmp6b-2,bmp6b-3,with full-length CDS of 1 245,1 257 bp,encoding 414 and 418 amino acids,respectively.Both bmp6a and bmp6b were unstable hydrophobic secretory proteins containing a signal peptide sequence and lacking a transmembrane structure.bmp6a was mainly distributed in mitochondria,nucleus and cytoplasm,while bmp6b was mainly distributed in nucleus and mitochondria.The secondary structure of bmp6a and bmp6b was mainly random coil,consistent with the results of tertiary structure prediction,and both contained one TGF-β-rel family conserved domain.The amino acid sequence homology analysis showed that the amino acid sequence of bmp6a had higher similarity with bmp6 of Megalobrama amblycephala,but lower similarity with mammals.The amino acid sequence of bmp6b had higher similarity with that of C.auratus and C.gibelio.Phylogenetic tree analysis showed that the bmp6a of C.auratus var.Qihe and the bmp6a of Cyprinus carpio were clustered in the same clade,while the bmp6b of C.auratus var.Qihe and the bmp6b of C.auratus and C.gibelio were clustered in the same clade.The results of qRT-PCR showed that bmp6a and bmp6b genes were expressed in brain,muscle,gill,spleen,intestine,ovary,liver and kidney tissues of C.auratus var.Qihe,but the expression levels were higher in liver,gill and kidney tissues.The expression patterns of bmp6a and bmp6b were similar in different developmental stages of juvenile C.auratus var.Qihe.The expression levels of bmp6a and bmp6b were the highest before the ossification of intermuscular spines,and then decreased with the development of intermuscular spines.To sum up,the CDS of bmp6 gene of C.auratus var.Qihe was obtained by cloning,and bioinformatics analysis and qRT-PCR detection were carried out,providing a reference for further creating a new strain of C.auratus var.Qihe without intermuscular bones.
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The aim of this study was to explore the effects and regulatory roles of HSP60 on the testes and ovaries,uterus and fallopian tubes before and after ovulation in Bactrian camels at different developmental stages.The CDS region sequence of the HSP60 was cloned from the Bactrian camel testis and its bioinformatics analysis was analyzed using ProParam and MEGA 7.0 software,and its expression was explored by polymerase chain reaction(PCR),hematoxylin and eosin(H&E)staining,immunohistochemistry(IHC),Real-time Quantitative PCR(qPCR)and Western Blot.The cloning results showed that the coding sequence(CDS)region of the HSP60 was 1 722 bp in length and encoded 537 amino acids,and the HSP60 sequence of the Bactrian camel was similar to that of dromedary camel and alpaca.The qPCR results showed that HSP60 was expressed in the testes of Bactrian camels at different developmental stages,and the expression levels in the testes after sexual maturity(3,5 and 7 years old)were significantly higher than that at 3 months of age.The mRNA expression of HSP60 was found in the ovaries,uterus and fallopian tubes before and after ovulation,and the mRNA levels in the ovaries and uterus were significantly higher after ovulation than before ovulation,but there was no significant difference in the fallopian tube before and after ovulation.Western Blot results showed that the expression trends of HSP60 in the testes at different developmental stages were similar to that of mRNA,and the protein expression level increased with age.The protein levels of HSP60 in the ovaries,uterus and fallopian tubes were significantly higher after ovulation than before ovulation.Immunohistochemical staining revealed that the HSP60 protein was primarily located in Sertoli cells,Leydig cells,part of spermatogenic cells,granulosa cells of the ovaries,glandular cells of the endometrium and muscle cells.The results showed that HSP60 was involved in testicular development and spermatogenesis in Bactrian camels,as well as in the regulatory processes of ovulation induction and meiosis.
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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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