As a key regulating factor in response to various abiotic stresses,plant heat shock transcription factor (Hsf) has a big family,and diverse structure,characteristics and functions.Hsf not only directly regulates Hsp and other relative gene expression and participates in the processes of response and adaption to various abiotic stresses,but also mediates many life activities regulation.Since the first Hsf was cloned from yeast in the 1980s,more and more Hsfs from other species have been identified and studied.In the previous reports,the identification of the Hsf family in plants was performed only in model species such as Arabidopsis and tomato.Furthermore,the studies is mainly focused on the HsfA subfamily,with few studies on the HsfB subfamily.And,the precise function of HsfC family is also largely unknown.With global climate change,the frequent occurrence of extremely high temperature events has seriously threatened the yield and quality of wheat,maize and other crops.To deal with the threat posed by heat stress,unraveling the mechanism of thermotolerance,identifying functional the targeted Hsfs and improving stress tolerance of crop through biotechnology methods is important.The number of Hsf family in field crops is various,the genome is complex,and the related research started lately compared with model species.To this end,our laboratory began to study the Hsf family of crops in 2009.Based on the latest genomic information,we confirmed the number of members,the modular structure and the spatio-temporal expression pattern of Hsf family.At the same time,with the help of transgenic wheat and mutant by genetic transformation and the CRISPR/Cas9 clustered regularly interspaced palindromic repeats (CRISPR)/CRISPR-associated protein mediated genome editing technology,several Hsfs were cloned and their regulatory functions of thermotolerance were identified,and some mechanism of thermotolerance was clarified.Our research not only enriched the theoretical basis of thermotolerance,but also provided new germplasm for biological breeding.At present,many studies have reported on functional identification and transcriptional regulation of Hsfs,however,evidences lack on which upstream component mediate Hsf's participation in regulation of thermotolerance,and the related mechanism is still unknown.Based on previous research results about wheat and maize Hsf families of lab,and many relative reports published in public,we reviewed the roles and mechanisms of plant Hsf in regulating process reported in recent years,aiming to promote research in illustrating the extensive and special roles and regulation network of plant Hsf family further,and dig useful genes and selective QTLs for biological breeding for plant thermotolerance.
01-0001-13_pic.png)
Light-harvesting chlorophyll a/b binding proteins are important in plant photosynthesis and abiotic stress response.To study the characteristics of GhLhcb2A1 and its expression patterns and functions in low temperature and drought response in upland cotton,full-length CDS of GhLhcb2A1 gene was cloned from the leaf cDNA of Jimian 262 by PCR.Bioinformatic analysis was conducted to learn the basic characteristics of the gene.The expression patterns and functions in low temperature and drought response were evaluated by qRT-PCR and virus-induced gene silencing.It was shown that the length of GhLhcb2A1 CDS was 798 bp,encoding 265 amino acids.GhLhcb2A1 was highly expressed in leaves and was significantly up-regulated in leaves and roots under low temperature and drought treatment.Compared with the control,its expression maximized at 3 h under low temperature and drought in leaves with 17.42 and 30.03 folds increase respectively,whereas maximized at 6 h under low temperature and 12 h under drought in roots with 11.65 and 65.04 folds respectively.Subcellular localization assay verified that GhLhcb2A1 was expressed in the chloroplasts of cells.Compared with the control plants,GhLhcb2A1 silenced plants showed a more severe phenotype of water loss and dryness under both low temperatures and drought.The accumulated malondialdehyde content in the leaves of the silenced plants was significantly higher than that of the control,while the proline content and superoxide dismutase activity were significantly lower than those of the controls,suggesting that GhLhcb2A1 silenced plant reduced the resistance to low temperature and drought.The above results implied that this gene played a positive role in regulating low temperature and drought response.
01-0014-08_pic.png)
Soybean P34 protein mainly exists in soybean seeds,and its upstream promoter was likely to regulate the high expression of downstream genes in seeds.In order to further study the tissue expression pattern of soybean P34 protein gene and the regulatory activity of soybean P34 protein gene promoter,qRT-PCR was used to detect the expression of soybean P34 protein gene in soybean tissues.The 5'upstream sequence of soybean P34 protein gene(GmP34P)was cloned.The transcription initiation sites and cis-elements were analyzed by bioinformatics.The expression vector was constructed and the tobacco was transformed by Agrobacterium-mediated leaf disk method to detect GUS expression in transgenic tobacco.The results showed that the expression of P34 protein gene in soybean seeds was significantly higher than that in roots,stems,leaves and flowers.The length of GmP34P sequence obtained by cloning was 1 380 bp.Predictive analysis showed that the transcriptional start site of this sequence was base A at position 1 342,and the sequence contained a variety of cis-acting elements related to high seed expression,such as RY element,Skn-1 motif,2S seed protbanapa,etc.The plant expression vector pCAM-GmP34P containing GUS gene driven by GmP34P promoter was obtained.The positive transgenic plants were screened by hygromycin,PCR and RT-PCR.The results showed that GUS gene expression was extremely significant in transgenic tobacco seeds compared with other tissues by qRT-PCR with positive pCAM-GmP34P transgenic tobacco plants.GUS histochemical staining showed that the GmP34P promoter could regulate the high expression of downstream GUS gene in seeds.
01-0022-07_pic.png)
WRKY is a unique class of transcription factors in plants,which plays an important role in plant abiotic stress response,seed dormancy and germination,growth and development,etc.In order to reveal the function and underlying molecular mechanism of GmWRKY44 gene in soybean WRKY transcription factor family,bioinformatics analysis and biology function verification of soybean Williams 82 GmWRKY44 were performed.GmWRKY44 gene was 1 077 bp in length and encoded 358 amino acids;the results of structural prediction and evolutionary analysis showed that,the secondary structure was composed of 23.46% α-helix,4.75% β-fold,58.94% irregular coil and 12.85% extended chain,and the tertiary structure was unified with the secondary structure;It contained a conserved WRKY domain, the zinc finger structure was of the C2H2 type, and it belonged to the WRKY IIc subfamily; GmWRKY44 is a homologous gene of Arabidopsis thaliana AtWRKY71 with a similarity of 35.56%, and the two genes had similar gene structures. RT-qPCR analysis showed that GmWRKY44 responded to salt stress and its expression level first decreased and then increased.Under salt stress,the germination rate and root length of wild-type(Col-0)and GmWRKY44 overexpressing Arabidopsis lines were inhibited to a certain extent,but GmWRKY44 overexpressing lines were significantly better than Col-0.In addition,under salt stress,the growth inhibition of GmWRKY44 overexpressing lines was lower than that of Col-0.Physiological index analysis revealed that under salt stress,the overexpression lines of GmWRKY44 exhibited significantly higher activities of superoxide dismutase(SOD),peroxidase(POD),and catalase(CAT)than Col-0,while the content of malondialdehyde(MDA)was significantly lower than Col-0.These data indicated that overexpression of GmWRKY44 could improve salt tolerance in transgenic Arabidopsis.
01-0029-07_pic.png)
In order to explore new approaches for high-oil peanut breeding and establish a new method for directly developing high-oil peanut germplasm,this study employed in vitro mutagenesis breeding technology to create new high-oil peanut germplasm.Jihua 9 embryo leaflet was used as mutagenic test materials,Jihua 9 and Jihua 54 were used as control test materials,and bleomycin was used as mutagenic agent.The ovules were sterilized and placed in gradient mutagenesis medium and screened for semi-lethal concentrations of bleomycin.After somatic embryos germinated into seedlings,sterile peanut seedlings were used as rootstocks,and transplanted to the field.Bioinformatics analysis of two known regulated peanut fat synthesis genes WRI1 and experimental feasibility validation by the correlation of WRI1 gene expression in grain and crude fat content of mutagenic plants were conducted.The results were best when the bleomycin was 3 mg/L.The crude fat content of IM13-3 was higher than that of Jihua 9(CK1,test variety control)and Jihua 54(CK2,high oil variety control).Two WRI1 genes,WRI1X2 and WRI1X1,encoding 366 and 357 amino acids,respectively,were both unstable hydrophilic proteins. WRI1 gene expression and crude fat content were significantly positively associated in grain.Bleomycin was first used as a peanut vitro mutagenesis agent,and IM13-3 was obtained with a crude fat content of 56.64%.It further proves the authenticity of Jihua 9 high oil mutant and the feasibility of peanut in vitro mutagenesis method. The gene expression level of the high-oil mutant WRI1 was determined and was significantly different from the control varieties. Demonstrate the feasibility of breeding methods for in vitro mutagenesis of peanut.
01-0036-09_pic.png)
To investigate the function of lysophosphatidicacid acyltransferase 2(LPAT2)in Brassica napus,from which one copy(A07)of BnaLPAT2 was cloned by PCR.we constructed the overexpression vectors p35S∷BnaLPAT2-A07 and the seed-specific expression vector pNapin∷BnaLPAT2-A07,and by utilizing Agrobacterium-mediated genetic transformation method,obtained a total of 15 and 11 transgenic Brassica napus cv.Zhongshuang 6 respectively by PCR positive detection.Real-time Quantitative PCR(qRT-PCR)showed that the transcript levels of BnaLPAT2-A07 in most tissues of T3 overexpressed rapeseed were higher than that of CK.However,in the seed-specific expression tissues of T3 transgenic rapeseed,the BnaLPAT2-A07 genes were strongly expressed in the development and maturation stages of silique.Soxhlet extraction results showed that the oil content in the transgenic seeds driven by the 35S or Napin promoter accumulated 1.39 and 2.36 percentage point more oil than control seeds,respectively.The fatty acid components of transgenic rape were detected by gas chromatography.Compared with CK,the content of linolenic acid was increased by 3.13 and 1.47 percentage point,respectively.Taken together,the BnaLPAT2-A07 could promote seed oil synthesis,however,the specific selection function of BnaLPAT2-A07 for linolenic acid needs to be further verified.
01-0045-08_pic.png)
The identification and expression pattern analysis of CmPIPs gene family in melon could provide theoretical basis and support for further exploring the function of CmPIPs gene family and the genetic improvement of melon.TBtools,MEME,MEGA X and Plant-CARE tools were used to analyze the bioinformatics of CmPIPs,and the expression level of CmPIP2;7 in the pericarp of melon at different stages after pollination,and the expression level of each member of CmPIPs in different tissues and different concentrations of plant hormone treated young leaves were visualized in the software GraphPad Prism 10.The results showed that CmPIP2;7 and CsPIP2;8 had nearest kinship;the 12 members of the CmPIPs family were mainly distributed on chromosomes 1,3,4,5,9,10 and 11;except that CmPIP2;8 had 3 CDS regions,the other members had 4 CDS regions.The promoter regions of each member of CmPIPs had multiple cis-acting elements,hormone responsive elements,such as auxin,gibberellin,and abscisic acid.The expression level of CmPIP2;7 was significantly up-regulated during rapid development and maturity of melon fruit.Members of various families of CmPIPs were expressed in different tissues of melon.After treatment with 40.0 μmol/L auxin,the expression level of CmPIP2;4 was significantly up-regulated,while the expression level of CmPIP1;1,CmPIP 2;1,CmPIP2;2 and CmPIP2;3 were extremely significantly down-regulated,and when the concentrations of abscisic acid were 0.4,4.0 and 40.0 μmol/L,the expression level of CmPIP1;1,CmPIP2;1,CmPIP2;3,CmPIP2;9 were significantly down-regulated.After treatment with 44.640 μmol/L methyl jasmonate,the expression level of CmPIP2;1 and CmPIP2;5 were significantly down-regulated,while the expression level of CmPIP2;2,CmPIP2;3,CmPIP2;7 and CmPIP2;9 were significantly up-regulated.When the concentration of ethylene glycol was 4.0 mmol/L,the expression level of each member of CmPIPs was significantly up-regulated.The gene structure,sequence characteristics,evolutionary relationship and collinearity of CmPIPs gene family members were clarified,and their expression pattern were analyzed.
01-0053-10_pic.png)
As one of the subfamily members of Ca2+-ATPase,ACA exerts a vital role in sustaining the intracellular Ca2+ concentration equilibrium and modulating plant growth and development in response to abiotic stress.To acquire an in-depth comprehension of the function of the lettuce ACA gene family,bioinformatics methodologies were utilized to identify and dissect the members of the lettuce ACA gene family.The outcomes manifested that 17 ACA genes were identified in lettuce,denominated as LsACA1 to LsACA17;the LsACA genes were unevenly distributed across 8 chromosomes;the subcellular localization prediction results disclosed that all LsACA proteins were localized in the plasma membrane;the number of introns exhibited significant variation(0—32)among the members of the LsACA gene family,a total of 15 conserved domains of LsACA proteins were identified,with amino acids ranging from 21 to 50;the proportion of the secondary structure was alpha helix>random coil>extended strand>beta turn;based on phylogenetic analysis,LsACA proteins were categorized into 5 subfamilies,namely Group Ⅰ to Group Ⅴ;according to collinearity analysis,it was found that there were fragment duplications in 6 pairs of genes,the Ka/Ks of their collinear gene pairs were all less than 1,signifying that purifying selection was the preponderant force in evolution.The expression patterns of LsACA gene family members under different calcium ion concentrations were analyzed by qRT-PCR.The results showed that:compared with the control,the expression levels of 12 LsACA genes in the calcium-sensitive variety Baoshilü under low calcium treatment were extremely significantly downregulated,while the expression levels of 9 LsACA genes in the calcium-insensitive variety Yeluo were extremely significantly upregulated and the expression level of 1 LsACA gene was significantly upregulated.The ACA gene family members of lettuce were identified and analyzed,and the characteristics of the LsACA gene family members were revealed.
01-0063-11_pic.png)
Chalcone synthase(CHS)is an important structural gene that regulates the early biosynthesis of flavonoid pathways in plants,and plays a role in plant growth and development and stress response.Previously,the key genes StCHS4 and StCHS5 for anthocyanin biosynthesis were identified in the potato CHS family by expression analysis.To further explore the function of potato StCHS4 and StCHS5 in the biosynthesis of flavonoids and anthocyanins,the characterization of StCHS4 and StCHS5 proteins was analyzed by online website.The 35S∷StCHS4-GFP and 35S∷StCHS5-GFP recombinant vectors were constructed by homologous recombination method based on the pRI101 binary vector,and then were transformed into Agrobacterium GV3101 strain.The subcellular localization of StCHS4 and StCHS5 proteins was determined by transient transformation of Nicotiana benthamiana.N.tabacum was used as the experimental material for transient overexpression and stable genetic transformation to analyze the content of total flavonoids and anthocyanins after overexpression of StCHS4 and StCHS5 genes.The results showed that the secondary structures of StCHS4 and StCHS5 proteins were mainly α-helix and random coil.StCHS4 was an unstable hydrophilic protein,and StCHS5 was a stable hydrophilic protein.The sequence alignment revealed that StCHS4 and StCHS5 had a close relationship with the CHSs of Capsicum annuum and Solanum lycopersicum,respectively.The results of subcellular localization revealed that StCHS4 and StCHS5 proteins were localized in the cytoplasm and cell membrane.In transient overexpression of tobacco,StCHS4 and StCHS5 genes significantly enhanced anthocyanin accumulation at 3—5 days after injection.Three transgenic N.tabacum lines of StCHS4 and StCHS5 gene were obtained,respectively.Compared with the wild type,the expression of StCHS4 and StCHS5 in transgenic plants was significantly higher,and the contents of total flavonoids and total anthocyanins were higher than those in the wild type.The total flavonoid content in StCHS4-OE3 and StCHS5-OE1 transgenic plants was significantly increased.The anthocyanin content in StCHS5-OE1 and StCHS5-OE2 plants increased by 89%,131%,respectively.The above results demonstrated that StCHS4 and StCHS5 were the key CHS genes in the flavonoid pathway of Solanum tuberosum,and the overexpression of StCHS4 and StCHS5 contributed to the biosynthesis of anthocyanins and flavonoids.
01-0074-10_pic.png)
DNA damage significantly hinders impact plant growth and development.NBS1 plays an important role in the cellular response to DNA damage,to analyze the functional difference between NBS1 and its alternative splicing isoform,NBS1-3.Specific primers were designed to clone NBS1 and NBS1-3 from the complementary DNA of Arabidopsis thaliana,and the sequences of the NBS1,NBS1-3 and the three-dimensional structure of the proteins were analyzed.Transgenic Arabidopsis thaliana lines overexpressing of NBS1 and NBS1-3 were generated respectively,and nbs1 homozygous mutant plants were identified,detecting the expression levels of NBS1 in overexpressed lines NBS1 and NBS1-3.To further elucidate the functional distinction between NBS1 and NBS1-3,wild type,mutant and overexpressed plants were treated with 0.6 mmol/L Methyl Methanesulfonate(MMS)and observed damage area.The results of quantitative showed that the expression levels of NBS1 in overexpressed lines NBS1 and NBS1-3 were higher than wild type.PI staining of root tips showed that after 0.6 mmol/L MMS treatment,the mutant showed the highest relative damage area,while the NBS1-3 overexpressed lines exhibited the least damage,followed by NBS1 overexpressed lines and wild type.These results suggest a potentially more important role of NBS1-3 in DNA damage repair compared to NBS1.
01-0084-09_pic.png)
To understand the member characteristics of the WRKY transcription factor family in hydrangea and their roles in response to leaf spot disease,this study utilized the bioinformatics method to characterize WRKY family members in Hydrangea macrophylla Endless Summer,and systematically analyzed the protein physicochemical features,gene structure,systematic evolution,collinearity and expression patterns under the infection with Corynespora cassiicola of WRKY family members.The results showed that there were 84 non-redundant HmWRKY members in H.macrophylla genome.All of the HmWRKYs were hydrophilic proteins and unevenly distributed on 18 chromosomes of H.macrophylla,encoding 112—1 046 amino acids.In addition,HmWRKY members were divided into 3 subgroups(Group Ⅰ—Group Ⅲ),which contained a conserved DNA binding domain composed of WRKYGQK and C2H2.The sequence length of HmWRKY varies greatly from 512 bp to 40 338 bp,and 8 collinear gene pairs with Ka/Ks ratios less than 1 were detected,indicating that the HmWRKY family experienced purification selection in evolution.18 HmWRKY members showed significantly differential expression after infection with C.cassiicola,of which 9 up-regulated and 9 down-regulated.The results indicated that these HmWRKY genes might play important roles in response to leaf spot disease of hydrangea.
01-0093-11_pic.png)
To reveal the sequence characteristics,subcellular localization,and expression pattern of PatASIL2,belonging to the Trihelix transcription factor family,the PatASIL2 gene was first cloned using the cDNA of Pogostemon cablin (patchouli)as a template,and the bioinformatics analysis was further performed.The PatASIL2-EGFP expressed vector was constructed and transformed into Arabidopsis protoplasm to investigate the subcellular localization of PatASIL2.The real-time quantitative reverse transcription PCR(qRT-PCR)was also performed to detect the expression profiles of PatASIL2 gene in different tissues of patchouli and under treatments of methyl jasmonate(MeJA),salt stress,drought stress,and cold stress.The results indicated that PatASIL2 gene contained an open reading frame of 1 035 bp and encoded 344 amino acids.The PatASIL2 protein was an unstable hydrophilic protein without transmembrane domains and signal peptides and had 41 serine phosphorylation sites and one Myb_DNA-bind_4 conserved domain.The phylogenetic analysis showed that PatASIL2 was classified into the SIP1 subfamily of Trihelix transcription factor family,and clustered closely with Sesamum indicum SiASIL2.The subcellular localization result indicated that PatASIL2 was a nuclear-localized protein.The qRT-PCR results indicated that PatASIL2 expressed in the young leaf,mature leaf,old leaf,stem,and root in patchouli,particularly with the highest expression in the old leaf.The PatASIL2 expression was significantly upregulated by MeJA at 12—24 h and salt stress at 3—24 h.The expression of PatASIL2 was significantly upregulated at 24 h after drought treatment,and at 12 h after cold treatment.
01-0104-09_pic.png)
Soil nitrogen(N)cycling enzyme activity serves as a crucial indicator for characterizing soil fertility and N transformation.To investigate the effects of long-term application of fertilizers on the soil enzyme activities correlated with N cycling in rhizosphere soil of double-cropping rice fields in southern China,our project was based on a continue 37-year fertilization localization field experiment,including four fertilization treatments:without fertilizer as a control(CK),single fertilizer(MF),rice straw residue and mineral fertilizer(RF),and 30% organic manure and 70% mineral fertilizer(OM).The activities of N cycling enzymes in the rhizosphere soil were measured,and their correlation with soil chemical properties was analyzed.The results were as follows:compared to MF and CK treatments,OM and RF treatments significantly increased the contents of total N(TN),organic carbon(SOC),ammonium N($\mathrm{NH}_{4}^{+}-\mathrm{N}$),nitrate N($\mathrm{NO}_{3}^{-}-\mathrm{N}$)and microbial biomass N(SMBN)in rhizosphere soil,and also increased rice yield.The urease(Ure)and nitrite reductase(NiR)activities of rhizosphere soil in OM and RF treatments were significantly higher than those in MF and CK treatments.The RF treatment significantly increased rhizosphere soil hydroxylamine reductase(HyR)activities compared to the other three treatments,by 21.7%,13.0%,and 8.7%,respectively.This finding shown that OM treatment significantly increased protease(Pro),nitrogenase(Nit),nitrate reductase(NR)and nitrous oxide reductase(Nos)in rhizosphere soil compared to RF,MF and CK treatments.In comparison to MF treatment,OM treatments increased Pro,Nit,NR and Nos activities in rhizosphere soil by 20.0%,26.1%,426.1% and 26.7%,respectively.Nonetheless,the activity of nitric oxide reductase(Nor)on rhizosphere soil was considerably higher in the CK treatment than in MF,RF and OM treatments.Pearson correlation analysis revealed a substantial positive correlation between soil NR,NiR,Nit,Nos,Ure,Pro and soil TN,SOC,$\mathrm{NH}_{4}^{+}-\mathrm{N}$,$\mathrm{NO}_{3}^{-}-\mathrm{N}$,SMBN as well as rice yield.Soil Nor activity was observed to have a significantly negative connection with soil TN,SOC,$\mathrm{NH}_{4}^{+}-\mathrm{N}$,$\mathrm{NO}_{3}^{-}-\mathrm{N}$,SMBN and rice yield.The findings presented above showed that soil chemical properties and yield were substantially related to rhizosphere soil N cycling enzyme activities.Redundancy analysis(RDA)showed that the first order axis could explain 93.34% of the enzyme activity in rhizosphere soil and soil $\mathrm{NO}_{3}^{-}-\mathrm{N}$,TN and SOC contents were the key factors affecting the pattern of rhizosphere soil enzyme activities.Therefore,the long-term application of organic materials such as organic manure and rice straw can enhance soil chemical and biological characteristics,stimulate soil N cycling enzyme activities,and effectively fertilize paddy soils by partially replacing chemical fertilizers.
01-0113-09_pic.png)
In order to provide production advice of high-quality upper tobacco leaves,field experiment was carried out to elucidate the effects of nitrogen application on the growth,development,carbon and nitrogen metabolism of upper leaves of different tobacco varieties.Three treatments,37.5(LN),75.0(MN)and 112.5 kg/ha nitrogen application rate(HN)were conducted with NC89 and Yunyan 87 as the test varieties.The effects of different treatments on the agronomic traits,photosynthetic characteristics,leaf tissue structure,key enzymes activity of carbon and nitrogen metabolism,and chemical composition of the upper leaves of were studied.The results showed that the leaf length,leaf width,leaf area,and leaf dry mass of the upper leaves of both varieties were significantly improved with the increased of nitrogen application rates.At 115 days after transplanting(DAT),the leaf area of NC89 and Yunyan 87 of HN were significantly increased by 63.10% and 68.43% compared to LN,respectively.Increasing nitrogen rate improved the leaf chlorophyll content of NC89,the leaf chlorophyll content of HN was 6.67%—37.50% higher than that of LN,respectively.The leaf net photosynthetic rate was notably improved by increasing nitrogen rate of Yunyan 87,especially at 70 DAT and 80 DAT.The leaf palisade tissue,leaf spongy tissue,and leaf thickness were markedly promoted with the increased nitrogen application rate on 85—115 DAT.The palisade tissue and spongy tissue thickness of LN and MN were almost stable from 95 to 115 DAT,but HN improved by 9.82%—14.08% and 10.72%—13.72% respectively.The leaf carbon content and leaf carbon and nitrogen ratio of both varieties were significantly decreased by reduced nitrogen rate,while leaf nitrogen content significantly increased.The activities of invertase,sucrose phosphate synthase,sucrose synthase,and glutamate synthase of both varieties were increased with the increased nitrogen application rate.HN decreased the amylase activity of Yunyan 87,but increased that of NC89.At 115 DAT,the amylase activity of HN of Yunyan 87 was 27.53% lower than MN,while HN increased amylase activity by 33.86% and 21.74% compared to LN and MN of NC89,respectively.The activity of glutamine synthetase of Yunyan 87 significantly increased with the increased nitrogen application rate,while there was no significant difference among different treatments of NC89.The content of flue-cured tobacco leaf reducing sugar and total sugar were reduced,but the content of nicotine and total nitrogen were improved with increased nitrogen application rate.Under the same nitrogen application rate,the content of flue-cured tobacco leaf nicotine,total nitrogen,and potassium of Yunyan 87 were higher than NC89,while the content of reducing sugar,content of total sugar(except LN),sugar and nicotine ratio,and nitrogen and nicotine ratio were lower than NC89.The effect of nitrogen application rate on upper leaves of tobacco varieties is difference.Increasing nitrogen application rate could promote the growth,development and carbon metabolism,reduce sugar and nicotine ratio and nitrogen and nicotine ratio,and improve the coordination of chemical components of NC89.However,increasing nitrogen application rate could lead to excessive nitrogen metabolism,delay the transformation from nitrogen metabolism to carbon accumulation metabolism,and cause late maturity of Yunyan 87.
01-0122-11_pic.png)
To clarify the regulatory effect of nitrogen(N)fertilizer reduction and postponing on the productivity of wheat-maize double cropping system in Huang-Huai-Hai Plain.The annual N fertilizer experiment of summer maize and winter wheat was established of four N application systems:annual N fertilizer application 400 kg/ha of traditional farmer treatment(F400),10% reduction of annual N fertilizer(FN),20% reduction of annual N fertilizer(FH),and 30% reduction of annual N fertilizer(FL)from 2020 to 2023 at Jiyang Experimental Base of the Shandong Academy of Agricultural Sciences in Jinan.The grain yield,aboveground N accumulation characteristics,N use efficiency,and the nitrate residue after harvest in the 0—200 cm soil layer of wheat-maize double cropping system were tested,in order to provide the theoretical basis for further optimization of N fertilizer management in Huang-Huai-Hai Plain.The results indicated that N fertilizer postponing was optimized the grain yield of summer maize and winter wheat under the condition of N reduction,and the averaged across the three years,FL significantly increased by 9.2%—18.1%,13.5%—20.5%,and 11.1%—19.1%,respectively,compared with F400 and FN.N fertilizer postponing improved the N accumulation rate,and promoted aboveground N accumulation at wheat-maize different growth stages,and the averaged across the three years,FL significantly increased plant N accumulation by 5.7%—12.3% and 5.0%—12.8% under silking and maturity,respectively,compared with F400,FN,and FH,as well as 8.2%—17.2% in grain N accumulation.For winter wheat,FL and FH treatments were significantly higher than F400 and FN at jointing,anthesis,and maturity,and the averaged across the three years,FL and FH significantly increased by 23.4%—28.1%,20.7%—26.3%,and 12.6%—20.8%,respectively,compared with F400,FN and FH,at the same time the grain N accumulation under FL significantly increased by 16.4%,15.0% and 5.8%,respectively,compared with F400 and FN.N fertilizer postponing optimized the N use efficiency of wheat-maize double crop system,the averaged across the three years,FL significantly increased N uptake efficiency by 4.8%—57.7% and 32.0%—72.4% of summer maize and winter wheat,respectively,compared with F400,FN,and FH;and FL significantly increased N partial factor productivity by 68.8% and 40.4% in summer maize,respectively,compared with F400 and FN,as well as by 38.4%—71.8% in winter wheat compared with F400,FN,and FH.At harvest of summer maize and winter wheat,the soil nitrate residue was mainly enrichment in the 0—40 cm soil layer under four N application systems,the averaged across the three years,accounted for 40.0%,38.9%,44.9%,42.5% and 37.3%,36.9%,46.7%,38.3% of the 0—200 cm soil layer,respectively.In addition,the obvious accumulated effects in 0—200 cm soil layer nitrate residue under F400 and FN treatments at harvest of summer maize and winter wheat,but there was the relative balance was achieved under FL and FH treatments.In conclusion,a 30% reduction of annual N fertilizer by N fertilizer postponing could optimize plant N accumulation characteristics and realized synergistic improve grain yield and N use efficiency.Therefore,FL treatment was an optimal N application system for realizing the collaborative target of high-yield,high-efficiency,and environment-friendly of wheat-maize double cropping system in Huang-Huai-Hai Plain.
01-0133-13_pic.png)
To explore the impact of organic fertilizer on the physiological growth of flax and the rhizosphere bacterial communities,and to investigate green high-yield cultivation techniques for flax in dryland,a field experiment was conducted using Baxuan No.3 as the material.The study examined the effects of four different fertilization treatments(T0:no application; T1:low quantity of cow manure; T2:medium cow manure; T3:high quantity of cow manure)on the physiological growth changes,nitrogen utilization,dry matter accumulation,and the diversity,community composition,co-occurrence networks,and metabolic pathways of the rhizosphere bacteria of flax,as well as discussing the environmental factors driving the differences in bacterial communities.The results showed that the T3 treatment resulted in higher flax production. Compared to the control, this fertilization condition also had the highest indicators for plant height, capsule fruit number per plant, thousand-grain weight, and nitrogen use efficiency, which form the physiological basis for stable yield following the application of organic fertilizer.The application of organic fertilizer significantly affected the diversity and richness of the rhizosphere soil bacteria of flax,and there were significant differences in the structure of the rhizosphere bacterial community.The population structure of the rhizosphere bacteria of flax was influenced by organic matter,total nitrogen,and available phosphorus.The dominant flora in the rhizosphere of flax was the same across different treatments,but the relative abundance of each dominant flora varied significantly.The rhizosphere of flax was dominated by the phyla Proteobacteria,Actinobacteria,Acidobacteria,Chloroflexi,and Bacteroidetes.The relative abundance of Proteobacteria and Actinobacteria increased with the increase of organic fertilizer treatments,while that of Acidobacteria decreased with the increase of organic fertilizer treatments.WGCNA analysis identified 15 co-expression modules,with the Red and Pink modules showing a significant positive correlation with organic matter content.The application of organic fertilizer increased the complexity of the bacterial network,and seven key OTUs were identified through combined WGCNA analysis.In conclusion,the application of organic fertilizer promoted the growth of flax and altered the structure and network complexity of the bacterial community in the rhizosphere soil of flax.
01-0146-11_pic.png)
In order to explore a good fertilization model to improve the fertility of yellow-cinnamon soil in Southern Henan,the characteristics of stable and high yield of wheat-maize rotation system and its relationship with soil nutrients under different fertilization treatments on yellow-cinnamon soil were studied.Based on the long-term experiment since 2012,four treatments were set up:no fertilizer(CK),chemical fertilizer(NPK),chemical fertilizer combined with manure(NPKM)and chemical fertilizer combined with straw(NPKS).Plants and soil samples were collected at maturity stage to determine crop yield and soil organic carbon,alkaline nitrogen,available phosphorus and available potassium.The results showed that compared with CK treatment,the crop yield of each fertilization treatment was significantly increased,the yield of wheat was increased by 53.70%—64.50%,and the yield of maize was increased by 44.54%—58.31%.The yield of NPKM treatment was the highest(wheat 8 162.61 kg/ha,maize 8 836.33 kg/ha),and there was no significant difference between NPKS and NPKM treatments.The yield sustainability index(SYI)of NPKM treatment was the highest,and the SYI values of wheat season and maize season were 0.84 and 0.82,respectively.The crop yield and its SYI value both showed NPKM>NPKS>NPK>CK,indicating that chemical fertilizer combined with organic materials could significantly increase crop yield and its sustainability.At the same time,different fertilization treatments could improve soil nutrients to varying degrees,among which NPKM treatment had the most significant improvement effect.The relationship between crop yield and soil nutrients was analyzed.The relationship between crop yield and soil nutrients was analyzed.There was significantly positive correlation between crop yield and soil organic carbon(SOC),available nitrogen and available phosphorus contents,among which the correlation between crop yield and SOC was the most significant.With the increase of soil organic carbon content,the SYI value of crops increased first and then stabilized,and the inflection point was 15.15 g/kg.To sum up,the application of chemical fertilizer combined with manure can significantly increase crop yield and soil nutrients,and maintain high crop yield sustainability.It is a recommended fertilization model to achieve sustainable production of soil-crop system in yellow-cinnamon soil ecological zone.
01-0157-09_pic.png)
The aim was to clarify the effects of different nitrogen application levels on soil organic nitrogen fractions and nitrogen use efficiency in maize fields in the central-western region of Inner Mongolia,so as to provide a reference for the scientific management of soil nitrogen and sustainable development of modern agriculture.Six nitrogen application levels were set up,N0(0 kg/ha),N8(120 kg/ha),N12(180 kg/ha),N16(240 kg/ha),N20(300 kg/ha),and N24(360 kg/ha).The dynamic changes with effects of nitrogen application on soil total nitrogen content,particulate organic nitrogen content,light fraction organic nitrogen content and heavy organic nitrogen content,as well as maize yield and nitrogen use efficiency were analyzed at different soil layers at pre-sowing and post-harvest.The results showed that soil total nitrogen,particulate organic nitrogen,light fraction organic nitrogen,and heavy fraction organic nitrogen content decreased with deepening of the soil layer at the same nitrogen application level;soil total nitrogen content at pre-sowing increased with the nitrogen application levels.Soil total nitrogen content in the N16,N20,and N24 treatments was significantly higher than that in the N0,N8,and N12 treatments at post-harvest.Soil particulate organic nitrogen content of N16 treatment was highest in the 0—10 cm,10—20 cm,and 20—40 cm soil layers at pre-sowing,with 0.14,0.13,and 0.09 g/kg,respectively.At post-harvest,N16 treatment had the highest content in the 10—20 cm,20—40 cm,and 40—60 cm soil layers,with 0.19,0.10,and 0.09 g/kg,respectively.The highest increase of soil light fraction organic nitrogen content of 37.27% was in the N16 treatment,and the highest increase of soil heavy fraction organic nitrogen content of 7.35% was in the N24 treatment,followed by the N16 treatment,at 6.84%.The N16 treatment had the highest maize biological yield of 31 443.50 kg/ha;the highest maize economic yield of 18 526.47 kg/ha;and the nitrogen use efficiency decreased with the increase in nitrogen fertilizer application levels,N16 treatment in the nitrogen harvest index was the highest, at 79.20%.In conclusion,the more suitable nitrogen fertilizer application level in the central-western region of Inner Mongolia should be maintained under 240 kg/ha,in order to achieve the best soil nitrogen management ang crop yield.
01-0166-11_pic.png)
To explore candidate genes for resistance of Aphis gossypii to the insecticide imidacloprid,the transcriptome data of the two strains were obtained and compared with imidacloprid indoor resistant and relatively sensitive lines of Aphis gossypii by using Illumina HiSeq 2500 high-throughput sequencing technology.The gene annotation was performed using the NCBI database,and bioinformatics analysis of the differential genes at the transcriptional level included GO function,KEGG metabolic pathway,and other analyses.The relative expression of eight candidate differentially expressed genes(CYP6a2,CYP6a13,CYP6k1,CYP6j1,CYP4c1,AChE2,CarE and ALP3)was detected using qRT-PCR technology,and the evolutionary relationships of resistance related genes were analyzed.After sequencing and sequence splicing,a total of 70 101 Unigenes were obtained,with an average length of 654.37 bp.29 131,27 861 and 2 993 Unigenes were annotated in NR,GO and KEGG databases,respectively.According to the NR annotation analysis of the differential genes of insecticide resistance and sensitivity strains,a total of 22 differential genes that may be related to insecticide resistance were found,including 9 detoxification enzyme genes(CYP6a13,CYP6k1,CYP6j1,CYP4c1 and ALP3 each,two CYP6a2,two CarE),8 cuticle protein genes(CP)and their precursors(CPP),target enzyme genes(AChE2),2 transcription factors(WRKY1,leucine zipper transcription factor-like protein 1 gene,LZTFL1),one pancreatic lipase-related protein 2 gene(PLRP2)and one multidrug resistance-associated protein gene(MRP).The results of qRT-PCR indicated that the expression levels of CYP6a2,CYP6a13,CYP6k1,CYP6j1,CarE and ALP3 genes in insecticide resistance strains were significantly higher than those in sensitive strains.The phylogenetic tree analysis of CYP,ALP,GST and CP genes obtained by NR annotation indicated that the genetic relationship between the Aphis gossypii and Aphis glycines and Acyrthosiphon pisum was relatively close.We found that 87.50% of the candidate differentially expressed genes in the two strains showed consistent changes in expression levels at the transcriptional and mRNA levels.
01-0177-12_pic.png)
The aim was to study the biological function of FolSid1 gene in Fusarium oxysporum f.sp.lini and its protein localization in Fusarium by cloning the gene.The gene sequence of FolSid1 was cloned by homologous comparison with F.oxysporum, and based on the principle of homologous recombination, a gene deletion box containing hydromycin resistance gene(hph)was constructed by Split Marker strategy,and the gene deletion mutant(ΔFolSid1)was obtained by PEG-mediated transfer into protoplasts of the wild type.pZESH1,a green fluorescent expression vector containing FolSid1 gene was constructed,and the subcellular localization of FolSid1-EGFP fusion protein was performed.The results showed that the sequence of FolSid1 gene consisted of 5 392 bp,which contained 3 introns.Compared with the wild type and the external insertion mutant, the knockout mutant ΔFolSid1 conidia showed a significant decrease in yield, although they did not differ in morphology and size; morphological observations revealed that the growth rate of colonies from the knockout mutant was significantly slower. The experiments of subcellular localization showed that FolSid1 protein was located in the cell membrane of mycelia cell.FolSid1 gene regulated the vegetative growth of mycelium,conidiogenesis and pathogenicity of Fusarium oxysporum f.sp.lini.
01-0189-10_pic.png)
In order to investigate the expression pattern and biological function of lncRNA TCONS_00143126 in E.coli type mastitis of cows in depth.This study used cDNA from bovine mammary epithelial cells as a template,and confirmed the presence of lncRNA TCONS_00143126 using PCR cloning and sequencing techniques.Subcellular localization analysis of lncRNA was performed,and potential target miRNAs and genes were predicted.The potential mechanism of its action in bovine mastitis was explored through KEGG pathway enrichment analysis.In addition,LPS was used to induce bMECs to construct an in vitro model of bovine mastitis,and the expression of lncRNA TCONS_00143126 in LPS-induced bMECs at 6,12 and 24 h was detected by RT-qPCR.The results showed that lncRNA TCONS_00143126 was real,and its expression was significantly up-regulated in LPS-induced bMECs,and it was mainly distributed in the nucleus.The results of target gene prediction and KEGG enrichment analysis showed that lncRNA TCONS_00143126 might regulate inflammatory signaling pathways such as JAK-STAT,mTOR and MAPK by targeting miRNAs(bta-miR-133a,bta-miR-193a-5p and bta-miR-375,etc.)and target genes(IFNE,SLC2A10,MEX3B),and then play a role in the inflammation of bovine mammary epithelial cells.
01-0199-08_pic.png)
Anoctamin 5(ANO5)is a multichannel membrane protein localized in the sarcoplasmic and sarcoplasmic reticulum that primarily plays a role in myosin membrane repair and phospholipid scrambling,mutations in the ANO5 gene can lead to jaw hypoplasia as well as various myopathies.It aimed to investigate the association of SNPs in the ANO5 gene with fat deposition traits in sheep.A population of 1 005 healthy and clearly genealogical Hu sheep male lambs was selected for the study,and PCR amplification and KASPar typing techniques were used to detect the locus polymorphisms of the ANO5 gene in the experimental population and analyze the associations with fat deposition traits.The expression level of ANO5 gene in different tissues was analyzed by qPCR.The results showed that sheep ANO5 gene was widely expressed in a variety of tissues in Hu sheep,and the highest expression of ANO5 gene was found in heart tissue compared with other tissues.Three genotypes of CC,CT and TT with the g.58010 C>T polymorphic locus were detected in the 10th intron of the sheep ANO5 gene.Descriptive statistics showed that the perirenal fat weight was the most different and had the highest degree of variability compared with other fat weights.Correlation analysis showed that fat deposition related traits were positively correlated with growth and feed efficiency traits,and the results of association analyses showed that the polymorphic locus was significantly associated with perirenal fat weight and its related traits in the Hu sheep.Among them,the perirenal fat weight of individuals with CC genotype was significantly lower than that of individuals with TT genotype.In conclusion,the g.58010 C>T mutation locus of the ANO5 gene can be used as a candidate molecular marker for perirenal fat deposition traits in Hu sheep.
01-0207-08_pic.png)
This study aimed to compare and analyze the milk composition,serum biochemical indices and expression levels of Leptin and LF genes during lactation between Bamei pigs and Yorkshire pigs.The six purebred Bamei pigs and Yorkshire pigs with healthy body conditions were selected respectively,which also had close parity,mating and delivery periods(within three days).A total of 50 mL milk,2 mL serum and 2 mL whole blood samples were collected from Bamei pigs and Yorkshire pigs on the 1st,7th,14th,21st and 28th day after delivery,respectively.Then,the milk,serum and whole blood samples were used to detect the milk compositions,serum indexes of alkaline phosphatase(ALP),albumin(ALB),globulin(GLB),triglyceride(TG),urea nitrogen(BUN),alanine aminotransferase(ALT)and aspartate aminotransferase(AST),and the expression levels of Leptin and LF genes.The results showed that on the 1st day of lactation in Baimei pigs,in addition to milk fat(Fat),the content of casein(CS),total protein(TP),total solid matter(TS),non-fat milk solids(SNF),lactose(L),free fatty acid(FFA)and acidity in the colostrum of Bamei pigs were all extremely significantly higher than those in the colostrum of Yorkshire pigs,while,the content of CS,TP,Fat,TS,SNF and L of Bamei pigs on the 21st day after delivery were extremely significantly lower than those of Yorkshire pigs.The serum biochemical indexes of two pig breeds showed different trends during the whole lactation.The content of ALT,AST,GLB,BUN and TG of Bamei pigs were significantly higher than those of Yorkshire pigs on the 1st day after delivery,while the content of ALP and ALB were all significantly lower than those of Yorkshire pigs.There were no different levels of these two genes on the 1st day,however,from the 7th day after delivery,the expression levels of LF in the Bamei pigs were significantly different than those in the Yorkshire pigs,while Leptin gene was extremely significantly expressed in Yorkshire pigs than in Bamei pigs.In general,the milk nutrients of Bamei and Yorkshire pigs showed a decreasing trend as the extension of the lactation periods,and the expression levels of Leptin and LF genes were likely related to the milk yield and quality between the two breeds,which may need to be further verified.
01-0215-09_pic.png)
The purpose of this study is to screen candidate genes related to the body weight at 8 weeks age of Kangle yellow chicken based on the omics data,and provide the theoretical foundation for molecular marker-assisted breeding of growth traits in Kangle yellow chickens.It also provides key basic data for improving molecular breeding methods for high-quality broilers and accelerating,the progress of breed selection.To detect SNPs significantly associated with body weight traits at 8 weeks age of Kangle yellow chickens,the body weight was measured from 8 to 22 weeks of age of 434 Kangle yellow chickens.Genome-wide association study(GWAS)was performed using the gene chip technology.Genes in the candidate regions with 2 Mb windows surrounding each significant SNP were found for GO and KEGG function analysis.Combined with the data of transcriptomic sequencing and published literature,key candidate genes related to the body weight at 8 weeks age of Kangle yellow chicken were screened.Two potential SNPs significantly associated with target traits were detected,located on chromosome 2(131 485 613 bp)and chromosome 4(60 413 848 bp).A total of 118 candidate genes were screened near SNP sites.Gene function annotation analysis showed the most significant enrichment of biological processes was retinoic acid metabolism.The significant enrichment of 12 KEGG pathways was found,including fatty acid degradation,tyrosine metabolism and drug metabolism-cytochrome P450.OXR1, RSPO2,EIF3E,TRHR,BMPR1B, ADH1C, MTTP, LAMTOR3, PPP3CA and PDLIM3 were preliminarily identified as key candidate genes for body weight traits at 8 weeks age of Kangle yellow chickens.Two SNPs and 10 key candidate genes were preliminarily identified related to the body weight at 8 weeks age of Kangle yellow chicken.
01-0224-08_pic.png)
In order to investigate the immune protective effect of apical membrane antigen 1(AMA1)of Eimeria tenella on chickens infected with E.tenella, the AMA1 gene amplified was connected to expression vector pET-32a(+).The recombinant plasmid was transformed into E.coli BL21(DE3)to express recombinant protein.108 chickens were randomly divided into 6 groups with 3 replicates in each group and 6 chicks in each replicate,including non-immune and non-attacking group(negative control group),non-immune attack group(positive control group),Freund's adjuvant group and 12.5,25.0,50.0 μg rEtAMA1 immunized groups.All of the chickens were injected with different concentration of recombinant protein at the age of 14,21 days,and infected orally with 5×104 sporulated oocysts of E.tenella at the age of 28 days and killed at the age of 35 days.The immune protection effect of rEtAMA1 was evaluated by body weight gain,lesion score,oocyst production,immune organ index and levels of cytokines and antibodies.There was no significant difference in average weight gain among the groups.Compared with the positive group,the scores of cecal lesions in 25.0 μg and 50.0 μg rEtAMA1 groups were significantly reduced by 53%,41%,the OPG in 12.5,25.0,50.0 μg rEtAMA1 groups were significantly reduced by 16%,27%,54% respectively,the immune organ index of bursa of Fabricius in 50 μg rEtAMA1 groups significantly increased by 27%,the immune organ index of thymus in 25.0 μg and 50.0 μg rEtAMA1 groups significantly increased by 25%,22% respectively.There was no significant difference in the immune organ index of spleen.Seven days after the second immunization,the levels of IL-2 and IFN-γ in 12.5 μg and 25.0 μg rEtAMA1 groups were significantly higher than the negative control group.The level of serum total IgG concentration in the 50.0 μg rEtAMA1 group significantly increased by 47% than that in the negative control group,and the concentration after the second immunization was obviously higher than that after the first immunization.The results showed that the rEtAMA1 could alleviate cecal lesions,decrease the oocyst production,promote the development of immune organs and increase the levels of cytokines and antibodies in serum,which had a certain immune protective effect on chickens infected with E.tenella.
01-0232-07_pic.png)
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
《Acta Agriculturae Boreali-Sinica》Official Website
Wechat Official Account
Copyright © 2022 《Acta Agriculturae Boreali-Sinica》
