To compare the effects of different nitrogen enhanced fertilizer on apple growth and to identify suitable nitrogen enhanced fertilizer and application methods for apple cultivation,this study examined the impact of four different types of nitrogen enhanced fertilizer on apple growth,yield,quality,and soil nitrogen supply capacity.The experiment consisted of seven treatments:no nitrogen fertilizer(CK),regular application of ordinary urea by farmers(U),coated urea mixed with ordinary urea 3∶7(CU1),basal application of coated urea and follow-up application of ordinary urea(CU2),loss-control urea(KSU),stabilizing urea(WDU),and humic acid stabilizing urea(FZU).A field experiment was conducted in Qixia City,Shandong Province,to analyze the effects of different fertilization treatments on the growth,yield,and quality of 6-year-old Golden Crown apple trees,as well as nitrogen utilization and soil nitrogen supply capacity.The results demonstrated that the application of diverse synergistic nitrogen fertilizers could markedly enhance spring and autumn growth,elevate leaf SPAD,and enhance apple yield and quality.Among these,the yield of the FZU treatment exhibited a significant increase of 8.89% in comparison to that of the control.The Vc content of the fruits of the CU1 treatment was found to be significantly increased by 66.73% in comparison to the U treatment.Furthermore,the CU1 treatment demonstrated the most pronounced impact on the improvement of growth in both spring and autumn.Nitrogen enhanced fertilizer had been demonstrated to significantly enhance nitrogen accumulation in new shoots and fruit nitrogen accumulation,nitrogen fertilizer agronomic efficiency,nitrogen fertilizer bias productivity and nitrogen utilization in the middle and late stages of apple fertility.Of these,the FZU treatment had been observed to exert the most pronounced effect.The nitrogen accumulation of new shoots in FZU treatment was significantly increased by 37.25%,15.91% and 37.85% compared with that in U treatment at bud differentiation,fruiting and ripening stages,respectively.The CU2 treatment was found to have the most beneficial effect on nitrogen utilization.Nitrogen enhanced fertilizer treatments demonstrated the capacity to significantly enhance the nitrate and ammonium content of soil.Among these treatments,the FZU treatment exhibited the most pronounced effect.The FZU treatment was observed to enhance soil urease activity during the flowering and bud differentiation stages.Additionally,the WDU and FZU treatments were found to significantly reduce the nitrification of soil ammonium and nitrogen loss.The {\mathrm{NO}}_3^{-}-N of the FZU treatment was concentrated at a depth of 20—60 cm soil,which was consistent with the distribution of apple roots in the soil and reduced the risk of {\mathrm{NO}}_3^{-}-N leaching.A comprehensive analysis demonstrated that the FZU treatment exhibited clear advantages in promoting apple growth,increasing apple yield and quality,and enhancing soil nutrient supply capacity.Consequently,the FZU treatment was identified as the optimal treatment.

To study the effect of different color bags on Marselan grape berry quality,and providing high quality raw material for vine culture.A complete randomized experiment design was used to analyze the effects of different color bags on berry quality(external quality and internal quality),peel quality and antioxidant activity.100 berries weight,reducing sugar,titrable acid and pH were increased in bags.The red bag increased the total phenols,total flavan-3-ols and total flavonoids,with 376.03,149.78,1 463.53 mg/kg,respectively,there were significantly differences for the total flavan-3-ols and CK,the total flavonoids and the CK.Blue and yellow bag treatments reduced the total phenols,total flavan-3-olsand total flavonoids.Bagging treatment significantly reduced total tannin content,the total tannin content of yellow bag was the lowest at 4.62 mg/g.Generally speaking,bagging treatment decreased DPPH free radical scavenging ability,FRAP(Reducing ability to Fe²⁺)and ABTS free radical scavenging ability,there was no significant difference for DPPH free radical scavenging ability and FRAP of red bag with CK,yellow bag significantly reduced FRAP,DPPH free radical scavenging ability and ABTS free radical scavenging ability.Correlation analysis showed that the soluble solids were positively correlated with reducing sugar and the sugar/acid ratio,the correlation coefficients were 0.98 and 0.95,respectively.In terms of peel quality,the total phenol was positively correlated with total flavan-3-ols and total flavonoids,the correlation coefficients were 0.95 and 0.98,there was a very significant positive correlation between total flavan-3-ols and total flavonoids,and the correlation coefficient was 0.98.For antioxidant activity,FRAP was very significant positively correlated with DPPH free radical scavenging ability and ABTS free radical scavenging ability free radical scavenging ability,the correlation coefficient were 0.74,0.86,respectively.Soluble solids,reducing sugars and sugar-acid ratio were significantly positively correlated with total anthocyanins,the correlation coefficients were 0.93,0.90 and 0.92,respectively.Principal component analysis was carried out on all the measured indicators,and two principal components with eigenvalues greater than 1 were extracted,the contribution rates of 78.31% and 10.09%,respectively,and cumulative contribution rates of 88.40%,which could basically represent most of the data of quality indicators.Red and blue bags were beneficial to berry quality,and the red bag was better,which can be used in Turpan region.

Fruit development is the key stage which determines the yield and quality of grape.WRKY family transcription factors play important roles in regulating plant development and environment adaptation.Ethylene is the important plant hormone which participates in regulation of fruit development,and ACC synthase is the key enzyme that limits ethylene synthesis.Using grape variety Zuoyouhong, VvWRKY13 overexpressing grape callus,as well as VvWRKY13 heterologous over-expressed tomato lines as materials,the role of VvWRKY13 from grape in fruit development and its relationship with ethylene were studied though plant physiological and biochemical methods as well as molecular biological techniques.The results displayed that the expressions of VvWRKY13 as well as ACC synthase genes VvACS2 and VvACS7 significantly upregulated at early stage of fruit development,and in VvWRKY13 over-expression grape callus,the expression of VvACS7C was significantly higher than control,but the expression of VvACS2 displayed no significant difference compared with control.Yeast one hybrid experiment showed that VvWRKY13 could directly bind to the VvACS7 promoter,VvWRKY13 had no direct interaction with VvACS2.We also found that the ethylene content and the expression of ACS family members,such as SlACS1b,SlACS4 and SlACS6,the ethylene synthesis genes,were significantly induced in VvWRKY13 heterologous over-expressed tomato,and the time from flowering to breaking of tomato was 3—6 days shorter than wild type;the above results indicated that,VvWRKY13 could regulate ethylene synthesis by promoting the expression of ACC synthase gene to participate in regulation of fruit development.

WRKY transcription factors play an important role in the regulation of plant growth and defence responses.In order to explore the function of WRKY gene family in chestnut under drought resistance,the chestnut WRKY gene family members were identified and the physical and chemical properties,phylogeny and structure of the encoded proteins were analyzed.Transcriptome sequencing and qRT-PCR were used to analyze the expression characteristics of WRKY gene family members under drought stress.The results showed that 65 WRKY gene family members were identified and divided into three groups:Ⅰ,Ⅱ and Ⅲ,among which group Ⅱ was divided into five subgroups according to its structure and phylogenetic relationship.Analysis of WRKY gene structure and conserved motif showed that the number of exons ranged from 1 to 7,and the number of exons and motif distribution in the same subgroup were similar.The conserved domain of WRKY gene had certain variation,including the deletion of WRKY heptapeptide domain,the deletion of zinc finger structure,and the heptapeptide domain was mutated into WRKYGKK,WRKYGRK and WRKYGRK.Transcriptome sequencing data showed that 4 WRKY family members were not expressed in the samples,and 49 WRKY family members were up-regulated under drought stress,while 11 WRKY family members were down-regulated under drought stress.The above results showed that,chestnut WRKY genes played an important role in response to drought stress.

To clarify the optimal harvesting period and storage temperature for the Venus apple fruit used for storage.The effects of three harvesting periods combined with three precise storage temperatures on fruit quality,anti-browning and preservation of Venus gold apple during 20 ℃ shelf life after cold storage were studied. The results showed that harvesting period Ⅰ(relatively early harvest)combined with storage at-1.5 ℃ (relatively low temperature)could maintain high fruit firmness,but the soluble solid content (SSC)of fruit was low,the taste and flavor of fruit were poor,and different degrees of browning of skin,heart and flesh appeared during shelf life. When harvested at period Ⅲ (relatively late),stored at 3 ℃ (relatively high temperature)until January 20,the fruit had high SSC,good taste and flavor. However,when stored at 20 ℃ for 24 h and stored on the shelf for 7 days,the fruit flesh was spongy and severely browning. The results showed that compared with-1.5 and 3.0 ℃,0 ℃ maintained higher firmness,SSC,titratable acid (TA)and ascorbic acid contents,and better maintained fruit chlorophyll fluorescence activity,and delayed fruit senescence. For the same storage temperature,the contents of TA and ascorbic acid in the fruits of harvest period Ⅱ were higher than those of harvest period Ⅰ and Ⅲ. The comprehensive analysis showed that the harvest period Ⅱ and Ⅲ combined with 0,3 ℃ could maintain the good merchantability and intrinsic quality of Venus gold apple stored in the medium and short term. For the Venus gold apple with medium and long-term storage,the harvest period Ⅱ combined with 0 ℃ accurate temperature storage could not only effectively inhibit fruit browning,slow fruit softening and senescence,but also maintain good flavor and quality of fruit during shelf life.

To find out the effect of laying reflective film on fruit quality in facilities and the mechanism of promoting coloring,the changes of the appearance and anthocyanin content of Jingyan grape fruits were monitored with the film laid between rows,then transcriptome sequencing and differential expression gene analysis of the peels with different coloring were carried out using high-throughput sequencing and Illumina Novaseq 6000 sequencing platform.Finally,41 151 986—46 673 908 Clean reads were obtained from 12 samples,with Q30 was over 94% and Q20 was over 98%.Besides,the mapping ratio between Clean reads and grape reference genome was 92.69%—94.22%.The analysis results of differentially expressed genes (DEGS) showed that there were 3 768 DEGs in CK 0 d—CK 7 d,with 1 488 up-regulated and 2 280 down-regulated.And 5 129 in RF 0 d—RF 7 d after film mulching,with 2 048 up-regulated and 3 081 down-regulated.It showed that film mulching can increase the differential expression of genes.Compared with CK,934 DEGs were obtained after 7 days mulching,of which 441 genes were up-regulated and 493 were down-regulated.KEGG enrichment analysis showed that,a total of 179 DEGs were annotated on 64 signal pathways,which were mainly enriched in five metabolic pathways:metabolism,cellular process,environmental information processing,genetic information processing and organismal systems.Particularly,many genes vere differentially expressed in benzene propane biosynthesis,endoplasmic reticulum protein processing,plant pathogen interaction and plant hormone signaling pathway.Results showed some transcripts of PAL,F3'H and 3GT were up-expressed,and some auxin-related protein genes were down-regulated.For transcription factors,WRKY gene was mainly up-regulated,while bHLH,C2H2,NAC,MIKC-MADS,Dof were down-regulated.In conclusion,laying reflective film may improve the activity of phenylalanine ammonia-lyase(PAL)and the expression of related genes(F3'H,3GT),thus promoting anthocyanin synthesis.Meanwhile,transcriptional factors WRKY,bHLH,C2H2,MYB,NAC,MIKC-MADS and Dof may also play an important role in regulation.

In order to explore the potential of melatonin to maintain the fruit quality of postharvest pear fruits,providing references for the research and development of storage and fresh preservation technologies of postharvest fruits,we carried out this research.In which Xinli No.7 pear fruits harvested at mature stage were selected as materials,and they were soaked in 0(control),0.02,0.20 and 1.00 mmol/L melatonin solutions for 2,4,8,12 and 24 h,respectively.Then we sampled and determined the effects of melatonin treatments on the internal qualities of pear,the activities of cell wall lysis related enzymes,the activities of antioxidant related enzymes,and the content of endogenous melatonin and salicylic acid in fruit peel and sarcocarp.The results showed that the hardness of the pear fruits soaked with 0.02,0.20 and 1.00 mmol/L melatonin for 8 h was 38.51,42.79,and 40.95 N/cm²,respectively,and the ethylene release rate decreased by 66%,70% and 71% respectively compared with the control.The soluble solid content(SSC)of pear fruits soaked with 0.2 mmol/L melatonin for 8 h was 94 g/L,which was significantly higher than that of the control(80.2 g/L)and was 1.17 times of the control.And the titrable acid(TAC)of pears of the same treatment was 1.013 g/L,which was also significantly higher than that of the control(0.90 g/L)and was 1.13 times of the control.In addition,treating pear fruits with exogenous melatonin could significantly increase the enzyme activities of superoxide dismutase(SOD),peroxidase(POD)and catalase(CAT),reduce the enzyme activities of polygalacturonase(PG),pectin methylesterase(PME)and Lipoxygenase(LOX),and could increase the endogenous melatonin content in fruit peel and the endogenous salicylic acid content in fruit(peel and sarcocarp).To sum up,soaking pear fruits with 0.2 mmol/L melatonin solution for 8 h had the maximum potential to maintain the fruit quality of postharvest pears,which could keep better fruit quality,keep lower cell wall lysis related enzyme activities and higher antioxidant enzyme activities,and could make the pear fruits having higher level of endogenous melatonin and salicylic acid content.

To provide a theoretical basis for the resource utilization of Chinese chestnut,the mature leaves of four chestnut cultivars were tested using ultra high performance liquid chromatography-tandem mass spectrometric,and functional secondary metabolites were screened by consulting relevant pharmacopoeias and searching the Chemical Book,Chemsrc,pharmacopeias and Web of Science database. The contents of 18 secondary metabolites we focused on in different cultivars were compared and analyzed. The study showed that a total of 673 kinds of secondary metabolites were detected in chestnut leaves,including flavonoids,phenolic acids,alkaloids,tannins,terpenoids,lignin and coumarin. Principal component analysis and cluster analysis showed that the difference of secondary metabolomics profile between Yanshanzaofeng and Yankui was little. The major secondary metabolites contributing to the classification were phloretin,didymin,salirepin,arbutin,cinnamic acid,gallocatechin,procyanidin B1 and so on. The differential metabolites could also be used as biomarkers to distinguish different chestnut varieties. A total of 202 functional secondary metabolites were selected,including 51 phenolic acids,24 alkaloids,83 flavonoids,21 terpenoids,14 lignins and coumarins,and 9 tannins. The contents of active substances were high in leaves such as protocatechuic acid,maslinic acid,ursolic acid,ellagic acid,chlorogenic acid,avicularin,kaempferol-7-O-glucoside,quercetin,spiraeoside,astragalin,betanin,tiliroside,ditymin and baimaside,which provided reference for the resource utilization of Chinese chestnut leaves.

In order to explore the effect of applying biochar on the improvement of soil physical and chemical properties in pear orchards and the continuous impact on the fruit quality of Huangguan pear,the positioning experiments for four consecutive years were conducted to provide theories and technologies support for rational and efficient fertilization in Huangguan pear orchards in Hebei Province.The experiment was carried out in the Huangguan pear experimental base in Jinzhou City,Hebei Province.Taking the treatment without biochar as the control(CK),the application of 0.45(B1),0.90(B2),1.35(B3),1.80 kg/m²(B4)4 treatments were used to study the effects of different amounts of biochar on soil organic matter,nitrate nitrogen,available phosphorus,available potassium,electrical conductivity and pH,and explored the effects on fruit quality(such as total soluble sugar,titratable acid,firmness,soluble solids,and single fruit weight)of Huangguan pear during ripening.The application of biochar for four consecutive years significantly reduced the soil nitrate nitrogen content,and increased the contents of organic matter,available phosphorus,available potassium and pH in the surface layer(0-20 cm),and the effect on the surface layer(0-20 cm)was greater than that in the 20-40 cm soil layer.B3 treatment had the best improvement effect on soil nutrients.The influence of soluble solid content on the fruit quality of Huangguan pear showed an increasing trend,and titrable acid showed a decreasing trend.There was no significant difference between B2 and B3 treatments on the quality of Huangguan pear.Considering the long-term cumulative effect of soil and the economic cost of biochar,0.90-1.35 kg/m² biochar application rate was recommended as the appropriate application rate.Combined with the analysis of the effects of the amount of biochar on the soil physicochemical properties and pear quality,under this test condition the recommended amount of biochar was 0.90 kg/m².

To reveal the soil factors that formed the quality characteristics of Hebei chestnut,54 typical chestnut orchards from 8 counties in Hebei Province were selected as the objects,and the contents of 9 mineral elements in the soil were studied by using significance,correlation and principal component analysis.The results showed that there were no significant differences in contents of Mg and Na,but there were significant differences in N,P,K,Ca,Fe,Mn and Cu contents in the soil of chestnut plantations in the main chest nut production areas.The N content in the soil of chestnut orchards was generally below the level of 3,and the P content was generally above the level of 3.The K content was generally at a low level,which was below the level of 5.The correlation of the contents of metal elements such as Ca,Mg,Fe,Mn,Cu and Na were significantly correlated with each other in soil.They were also significantly correlated with P element,and the correlation with N content was generally weak.Principal component analysis showed that mineral elements in the first 3 principal components reflect the total information content 73.521%.The contribution rate of the first principal component accounted for 44.678%,and the elements which included Ca(0.871),Cu(0.840),Fe(0.809)and Mg(0.806)were the main factors.It showed that the metal trace element content contributes more to the principal component.The comprehensive score was based on the mineral elements content of different chestnut orchards,which decided the order of high and low.The highest score was in Qianxi County(0.591 8).The ranking of the soil quality in the main chest nut production areas was:Qianxi>Qinglong>Xinglong>Kuancheng>Shahe>Zunhua>Xindu>Funing.To sum up,it is confirmed that the comprehensive scores of metal elements contents such as Ca,Mg,Fe,Cu can be used to evaluate the soil quality by using principal component analysis.It was advisable to select high K and low P compound fertilizer,and it was recommended to apply multiple elements mixed fertilization as a supplementary measure to improve the soil quality level of chestnut orchards.

Cold resistance is an important target trait for peach breeding and is regulated by multiple genes.Transcriptome sequencing is used to sequence transcripts under specific physiological conditions to explore the molecular mechanism of low temperature response of peach leaves.Leaves of Xiongyuejutao under 4 ℃ low temperature(LT)treatment and normal leaves(RT)were analyzed by transcriptome sequencing to explore the molecular regulation mechanism of cold resistance.A total of 72 up-regulated differentially expressed genes and 3 down-regulated genes were obtained by DESeq2.KEGG analysis was used to analyze the functions of differentially expressed genes in MAPK signaling pathway and plant-pathogen interaction pathway,and protein interaction was predicted.The results showed that leaves under low temperature treatment showed physiological states of curled,yellow,and water loss.The differentially expressed genes were mainly enriched in plant-pathogen interaction,MAPK signaling pathway,and secondary biomass accumulation.MYC2,SPCH and Pti4—6 transcription factors may play an important role in the regulation of cold-induced injury in response to cold stress.MYC2 and CBF proteins were predicted to interact with each other and interacted with multiple proteins.The results showed that multiple metabolic pathways responded to low temperature stress in peach leaves,and related transcription factors that may play a key role in the regulation of cold response pathways were predicted,which provided research direction and basis for the exploration of molecular mechanism of cold response regulation in peach.

In order to explore the effects of amino acid water-soluble fertilizer on grape yield and quality,the full-fruit Zaohebao grape was used as the experimental material,three treatments were set up:conventional fertilization (CK);increased application of organic fertilizer (OPT);organic fertilizer + amino acid water soluble fertilizer treatment (OPT+AA),the effects of exogenous amino acids on grape yield,growth,quality and soil nutrient content were studied.The results showed that compared with CK,grape yield decreased by 14.17% under OPT treatment and 7.50% under OPT+AA treatment,but were not significant.Compared with CK,partial productivity of fertilizer treated with OPT and OPT+AA increased significantly.Partial productivity of N,P₂O₅ and K₂O treated with OPT+AA increased significantly by 73.44%,85.01% and 85.01%,respectively.OPT+AA significantly increased total sugar by 8.09%,soluble solids by 1.32 percentage,sugar-acid ratio by 38.80%,and the titratable acid content by 0.16 percentage.The nutrient content of shoots was significantly increased by 24.73% in OPT+AA,total phosphorus content and total potassium content were significantly increased by 24.73%.In addition,OPT and OPT+AA treatments did not inhibit plant growth,but could effectively reduce the leaching of available nutrients and increase the nutrient content in the root domain.In summary,reducing and increasing the application of organic fertilizer had no significant effect on grape yield,but could reduce the leaching of available nutrients and significantly improve the partial productivity of fertilizer.The application of amino acid water-soluble fertilizer could promote nutrient absorption in plants,improving grape yield and fruit quality.

In order to study the function of 3-O-glucosyltransferase gene(3GT) in blueberry and to explore the roles of 3-O-glucosyltransferase in anthocyanin synthesis, a single copy of 3GT was cloned from the cDNA of blueberry fruit by RT-PCR, which was 1 371 bp and encoded 456 amino acids. 3GT was located at the end of chromosome Ⅳ in blueberry and contains 3 exons and 2 introns. The molecular formula of the protein was C₂₂₇₉H₃₅₂₁N₅₉₅O₆₅₀S₁₅, and the theoretical isoelectric point (pI) was 6.14. Information analysis predicted that 3GT was one hydrophilic protein and unstable protein with 37 phosphorylation sites, without signal peptide,or transmembrane domain. The secondary structure of protein consists of α-helix (Hh), extended chain (Ee), β-angle (Tt) and random curl (Cc), among which α-helix accounted for 40%.As one number of GTB-type glycosyltransferase family, the 44th position in the PSPG domain of 3GT was histidine, and the glycosylase donor might be UDP-galactosewas. 3GT was closely related to Kiwifruit, with the homology of amino acids reached 67%. Auto-Dock showed that 3GT had four interaction forces with zymolyte of cyanidin. There were multiple cis-regulatory elements that might relate to light response,stress resistance response and specific hormone response in the promoter of 3GT. The blueberry 3GT gene was cloned.The bioinformatics analysis and function prediction were carried out through online website and prediction software to study the characteristics and function of 3GT.

In order to investigate the effect of 1-methylcyclopropene(1-MCP)on the physiology,storage quality and electronic nose characteristic of Starking delicious apple.Fruits were treated with 1-MCP and then stored at ambient temperature(20±1)℃.Respiration rate,ethylene production rate,fruit intrinsic quality,color difference and the electronic nose response was analyzed during storage.The results showed that the postharvest respiration rate and ethylene release rate of Starking delicious apple increased at first,then decreased after peaking at 10 d and 15 d,respectively.At the same time,fruit firmness and titratable acid(TA)content decreased,soluble solid content(SSC)increased then decreased.Fruit chromaticity varied,a^*,b^*,C^* and ΔE^* values increased.1-MCP treatment inhibited fruit respiration rate and ethylene release rate,delayed the decline of fruit firmness,SSC and TA content,inhibited increasing of a^*,b^*,C^* and ΔE^*.1-MCP treatment significantly reduced the production of sulfides and terpene compounds(W1W),nitrogen oxides(W5S),organic sulfides and aromatic compounds(W2W),methyl aromatic compounds(W1S),alcohols,aldehydes and ketones aromatic compounds(W2S).Linear discriminant analysis(LDA)combined with electronic nose could distinguish control and 1-MCP fruit at different storage periods.Loading analysis showed that W1W,W5S,W2W,W1S and W2S sensors played key roles in differentiating control and 1-MCP treatment fruit during storage.During ambient temperature storage,1-MCP treatment delayed fruit softening and kept a lower solid-acid ratio,effectively maintained fruit flavor and color of Starking delicious apple.However,1-MCP decreased the response value of sensitive sensors in electronic nose,and inhibited the production of fruit volatile substances.According to correlation analysis,electronic nose sensor response values were significantly correlated with the internal quality and color difference,which may provide a basis for the rapid and nondestructive detection of Starking delicious apple.

In order to clarify the biological function of gibberellin receptor GID1 in Pyrus betulifolia,and provide a good foundation for future development of P.betulifolia dwarf rootstocks using CRISPR/Cas9 gene editing technology.Pyrus betulifolia was used as the test material,and the PbGID1s genes were obtained by homologous cloning method.Bioinformatics analysis software was used to construct the gene structure and design the target sites;construction of sgRNA expression cassettes with targets into CRISPR/Cas9 expression vectors,through the mediation of Agrobacterium,the CRISPR/Cas9 expression vector was transferred into the cotyledons of P.betulifolia.Results showed that four PbGID1s were successfully cloned from P.betulifolia plants and named as PbGID1b-1,PbGID1b-2,PbGID1c-1 and PbGID1c-2. They all consisted of two exons and one intron found by gene structure analysis.Amino acid sequence comparison showed that all PbGID1s had the HGG and GXSXG conserved domains.Five gRNAs that could potentially edit all 4 PbGID1s simultaneously were successfully constructed into a single CRISPR/Cas9 vector,pYLCRISPR/Cas9P_35S-N.The results of the genetic transformation test of P.betulifolia showed that a total of 595 cotyledons of P.betulifolia were infiltrated,176 resistant buds and 33 positive plantlets were obtained,and the transformation efficiency reached 5.55%.A CRISPR/Cas9 vector was successfully constructed that could simultaneously target the PbGID1s family genes of P.betulifolia.Through the mediation of Agrobacterium,the vector was successfully transformed into P.betulifolia cotyledons,and positive plants were obtained.

Strawberry powdery mildew caused by Sphaerotheca aphanis is one of the strawberry diseases with large incidence area and high incidence frequency in the protected cultivation areas, which usually occurs from seedling stage to fruiting stage. Therefore, the establishment of a rapid and efficient detection method of S. aphanis is quite important for the diagnosis and control of strawberry powdery mildew.A set of LAMP amplification primers including F3/B3、FIP/BIP and LF/LB were designed for the conserved sequence of ITS gene of Sphaerotheca aphanis as the target gene. Through the double judgment method of Real-time fluorescence curve and fluorescence color change, the main factors in the LAMP reaction system including the final concentrations of dNTPs, MgSO₄, BstDNA polymerase and betaine and amplification temperature were selected and optimized to increased etections ensitivity and specificity. Finally, the effects of field samples detection by optimized LAMP were evaluated.The results showed that the LAMP detection system of strawberry powdery mildew was established successfully. The optimized reaction conditions included dNTPs 1.6 mmol/L, MgSO₄ 8 mmol/L, BstDNA polymerase 320 U/moL and betaine 1.2 mmol/L and the amplification temperature was 65 ℃. Under such conditions, the minimum detection sensitivity was 3.2×10^-4 ng/μL within 60 min, and the efficiency of which was 100 times higher than the result of PCR amplification.The visual LAMP method could effectively detect the samples with no obvious symptoms infected by S. aphanis in the early stage. This method has the advantages of short detection time, direct observation with eyes, low requirements for operators and low detection cost. It has important guiding significance for the early diagnosis, monitoring, early prevention and determination of the best control time of strawberry powdery mildew.

Grapes are often subjected to various abiotic stress during their growth and development,such as drought,salt,heat or chilling,among them,drought stress is one of the most important abiotic stresses inhibiting vegetative and reproductive growth of grape.Although grapes are more resistant to drought stress,about half of our country's main grape cultivation area belongs to arid and semi-arid climates,drought is one of the main factors restricting the development of our country's grape industry.To ensure the healthy development of our country's grape industry,the research on the effects of drought stress on grape,the formulation of rational irrigation system and the breeding of drought-resistant varieties have become the research hotspots in recent years.Due to the extensive effects of drought stress on grape,in older to cope with drought stress,grapes have evolved many regulatory mechanisms to balance the effects of drought stress in grape growth.Firstly,the water demand degree of grape in different growth period was analyzed according to the law of grape water consumption,and the effects of drought stress on main physical and chemical indexes were exhibited,including photosynthesis,osmotic regulation and reactive oxygen regulation.And then,the quality and yield of grape fruit were analyzed under drought stress,the effects of drought stress on grape fruit quality were reviewed.We give the prospect for how to use drought stress strategy to improve grape quality and how to further study the molecular mechanism of grape response to drought stress at last.

In order to explore the storability of Chinese cherry fruit,reduce the decay rate of post harvest fruit and prolong its storage life. Chinese cherry Black pearl was sprayed with CaCl₂,amino acid calcium and sugar alcohol calcium after flowering. The internal and external quality of post harvest fruits during storage and several protective enzymes were studied,and the most suitable calcium fertilizer and its concentration were selected.The results showed that:with the extension of storage time,cherry fruit hardness,100 fruit weight,soluble sugar,titratable acid,soluble protein,Vc,β-carotene decreased;the content of MDA and REC increased gradually;the activities of soluble solids,SOD,POD and PPO increased first and then decreased;the nutrient and water loss,PPO activity and MDA accumulation of cherry fruit treated with calcium fertilizer during storage were lower than CK;the activities of SOD and POD were higher than CK. Foliar calcium fertilizer improved the storage resistance of Black pearl cherry. The comprehensive evaluation of membership function showed that 1 500 times sugar alcohol calcium had the best effect on fruit preservation. Spraying calcium fertilizer before harvest could improve the hardness,quality and the activity of antioxidant enzymes of cherry fruit,so as to prolong the storage period of cherry.1 500 times sugar alcohol calcium can be popularized and applied in production.

In order to explore the effect of RNA extraction and virus detection of different parts of apple organs,apple plants infected by three viruses were used as experimental materials.Adsorption column method was used to extract total RNA from flowers(pistil,anther,filament,petals,calyx,receptacle,and flower stalk),leaves(petiole,mesophyll,and veins),fruits(fruit stalk,peel,flesh,score,episperm,and endosperm)and branches(phloem),a total of 17 tissues of four organs.The extraction effects of different sampling parts were compared,the detection effects of three viruses in different parts of organs were analyzed by regular RT-PCR,and the relative content of viruses in different parts of organs were analyzed by Real-time fluorescence quantitative PCR.The results showed that the effects of RNA extraction from different organs of apple were different,with the highest concentration in leaves,followed by flowers and phloem of branches,and the lowest concentration in fruits.Anther,filament and petiole were best extracted from flowers and endosperm and petiole were best extracted from fruits and leaves,respectively.Three apple viruses could be detected in 16 tissues except endosperm.Quantitative analysis showed that the relative content of the three apple viruses was the highest in flowers.Furthermore,the relative contents of viruses in pistil,episperm and petiole were correspondingly higher than that in other tissue parts of flower,fruit and leaf,respectively.In addition,the content of the three viruses in different tissue parts of fruits and leaves showed similar trends,but there were certain differences in the distribution of different tissue parts of flowers.

In order to determine the effects of V.villosa Roth cultivation in apple orchard on soil microbial community structure,the high-throughput sequencing technology was used to analyze the microbial structure of V.villosa Roth cultivation soil in seven areas of Shandong Province and different phenological period of the same orchard,taking natural grass treatment as comparison.The correlation between microbial community structure and physicochemical properties of grass cultivation soil was analyzed.Results showed that,in seven sampled sites of Shandong Province,the relative abundances of Metarhizium in V.villosa Roth cultivation treatment were all higher than those in the control,and the difference was biggest in Dongying Area.The relative abundances of Fusariumin V.villosa Roth cultivation treatment of six sites were lower than those of the control,except for Zoucheng.The relative abundances of Gibberella in V.villosa Roth cultivation treatment of six sites were lower than those in the control,except for Rongcheng.In Zoucheng and Longkou,the relative abundances of Alternaria were lower than those of the control.Analysis of microbial community structure in different apple phenological stages of V.villosa Roth cultivation in the same orchard showed that,the relative abundance of Aspergillus in flowering stage soil was much lower than that of natural grass.The relative abundances of Fusarium and Aspergillus in fruit setting stage in V.villosa Roth cultivation treatment were lower than those of natural grass.The relative abundance of Bacillus in V.villosa Roth treatment was higher in fruit expansion stage.The relative abundance of Bradyrhizobium was higher in mature stage,while Gibberella,Cladosporium and Bipolaris were much lower than those of natural grass.Effects of V.villosa Roth cultivation in apple orchard on soil microbial community structure differed in different region and phenological stage.However,the relative abundance of beneficial bacteria in soil was increased,while that of pathogenic fungi was decreased,which was helpful to improve orchard soil micro-ecological environment and promote the healthy cultivation of fruit trees.

To clarify the relationship between BcPDR1 gene and MAPK pathway genes BcBMP1 and BcBMP3 of Botrytis cinerea.We analyzed the expression levels of BcBMP1 and BcBMP3 in mutants of BcPDR1 by Real-time PCR and the expression levels of BcPDR1 in RNAi mutants of BcBMP1 and BcBMP3.In the background of mutant ΔBcpdr1,strains ΔBcpdr1/BcBMP1-OE and ΔBcpdr1/BcBMP3-OE over-expressing BcBMP1 and BcBMP3 were constructed,and the phenotype and pathogenicity of the over-expressing strains were analyzed.It was found that the expression levels of BcBMP1 and BcBMP3 in BCt89 and ΔBcpdr1,gene mutants of BcPDR1,were significantly better than those of the wild-type strain and the revertant strain,and the expression levels of BcPDR1 in the RNAi mutants of BcBMP1 and BcBMP3 gene were obviously lower than those of the wild-type strain.The colony morphology,hyphal morphology,growth rate,sporulation,and pathogenicity of the over-expression isolates ΔBcpdr1/BcBMP1-OE and ΔBcpdr1/BcBMP3-OE were obviously different from ΔBcpdr1,and closer to the wild-type BC22.These results indicated that the BcPDR1 was closely related to the expression levels of BcBMP1 and BcBMP3 in B.cinerea.The BcPDR1 gene negatively regulates the expression of BcBMP1 and BcBMP3,while BcBMP1 and BcBMP3 genes positively regulate the expression of the BcPDR1.

In order to explore the mechanism of grape defense response to grape downy mildew,the leaves of Muscat and Moldova were inoculated with grape downy mildew in vitro and cultured in light incubator.The development of downy mildews was observed from macroscopic and microscopic aspect.In macroscopic,it was showed that white mildew layer appeared on the back of leaves of Muscat four days after inoculation,and the area of disease spots expanded with time,meanwhile chlorosis yellow spots appeared on the opposite position of the front side of leaves.However,there were only a few black spots on leaves of Moldova at the inoculation point.Under electron microscope,it was found that the spores of the pathogen invaded the stomata and colonized,and germinated new sporangium stalks,which released the spores and propagated on the leaves of Muscat.But the number of sporangia was small on the leaves of Moldova,and most of the stomata was closed,only with spores,hyphae and tissue fragments scattered around the stomata.Furthermore,transcriptome sequencing and differentially expressed genes were analyzed by Illumina high-throughput sequencing.A total of 664.86 Mb of high-quality reads were obtained by RNA-seq.GO enrichment analysis showed that differentially expressed genes were mainly concentrated in biological pathways(metabolic process,cellular process,response to stimulus and biological regulation category),cell components(cell, membrane, membrane part and organelle),molecular functions(catalytic activity and binding,transporter activity and transcription regulator activity).KEGG enrichment analysis showed that the differentially expressed genes were mainly enriched in the plant pathogen interaction pathway and MAPK signal transduction pathway.Nine genes that may be related to grape disease resistance were screened and verified by Real time PCR analysis,and the results were consistent with the transcriptome sequencing results.

To provide a theoretical basis for the high-quality production and precise fertilization of Merlot grapes,through the determination of the mineral element content of each growth and different tissue parts,and the nutritional diagnosis and analysis based on the quality index formed by the phenolic substances in the fruit.The comprehensive quality index of fruit phenolics,namely CI value,was calculated by Topsis analysis method.By comparing the correlation between content of various mineral elements in different growth stages/ tissues and CI value,the nutritional diagnostic factors are determined.In order to provide a theoretical basis for the high-quality production and precise fertilization of Merlot grapes,high-CI vineyards were classified by CND method.The results showed that there was a significant synergistic and antagonistic relationship among the mineral elements in the leaves,petioles and fruits of the Merlot grapes.VS_F_N,MS_F_P,VS_F_K,VS_L_Ca,FBS_F_Mg,VS_F_Fe,FBS_L_Mn,MS_F_Zn,FBS_P_Cu,VS_P_B,FBS_L_Mo were selected as plant nutrition diagnostic factors.The quality index inflection point value of high-CI vineyards was 0.735 5,of which 7 orchards met this condition,accounting for 14.58% of the overall sample.According to the mineral element content range of Merlot plant within high-CI vineyards,the appropriate values of nutritional diagnostic factors were:N(8.85—11.81)mg/g,P(1.98—4.26)mg/g,K(14.97—20.70)mg/g,Ca(35.57—68.83)mg/g,Mg(3.69—15.51)mg/g,Fe(70.96—103.26)mg/kg,Mn(166.20—277.67)mg/kg,Zn(10.71—20.27)mg/kg,Cu(9.54—14.90)mg/kg,B(11.44—17.07)mg/kg,Mo(0.69—1.60)mg/kg.The nutritional diagnosis of high-CI vineyards showed that the K,Ca,Mn,and Mg in Merlot plants were on the low side.The obtain the highest fruit quality index as the goal,the recommended fertilizer rates per hectare were N 62.25 kg,P₂O₅ 46.50 kg,K₂O 0.00 kg,CaO 56.25 kg,and MgO 46.50 kg.Ca and Mg fertilizers should be applied in small amounts and multiple times;we should strengthen the management of water and fertilizer in the orchard to increase the absorption of K element by the tree.Through foliar spraying,the lack of Mn elements can be appropriately supplemented.

In order to explore the mechanism of AcWRKY70 transcription factor in response to kiwifruit canker stress in different resistant varieties,AcWRKY70 gene was cloned from the leaf cDNA of resistant variety Xuxiang and highly susceptible variety Hongyang kiwifruit.The sequence structure characteristics of Hongyang AcWRKY70 gene,subcellular localization and evolutionary relationship of Hongyang AcWRKY70 protein were analyzed. The expression patterns of AcWRKY70 gene,including the tissue expression and different expression in resistant kiwifruit varietie and susceptible variety under Pseudomonas syringae pv. Actinidiae(Psa),salicylic acid(SA)and methyl jasmonate(MeJA)treatment were analyzed by RT-qPCR. Results showed that the full length of the Hongyang AcWRKY70 gene was 906 bp(GenBank accession number was MW881147). AcWRKY70 gene contained 885 bp of open reading frame(ORF)and encoded 294 amino acids. AcWRKY70 protein had typical WRKYGQK domain and the zinc finger structure was C2-HC. It belonged to class Ⅲ group of WRKY family and islocated in the nucleus. Phylogenetic tree analysis showed that it had the closest genetic relationship with tea plant. Both Hongyang and Xuxiang AcWRKY70 genes had the highest expression level in leaves. Under Psa treatment, the highest expression of AcWRKY70 gene reached at 12 h in Xuxiang, the expression of Hongyang reached the maximum at 48 h.Under the co-treatment of salicylic acid and Psa(SA+Psa),Methyl jasmonate and Psa(MeJA+Psa),the highest expression of AcWRKY70 gene reached at 12 h in Xuxiang. Under SA+Psa and MeJA+Psa treatment,the expression of Hongyang reached the maximum at 72,24 h,respectively. AcWRKY70 gene played a certain role in resistance stress of kiwifruit,and the response mechanism to pathogens in different resistance kiwifruit varieties may vary considerably.

Discuss the difference and change rule of texture characteristics of Huahong and Huayue apples after different shelf temperature combined with 1-MCP treatment during the shelf life,in order to determine the appropriate shelf temperature and time for apples.Using Huahong and Huayue as the test materials,the physical property analyzer texture multi-faceted analysis(TPA)and puncture test were used to determine the pulp elasticity,gummy stickiness, chewability, pulp hardness, rupture work, rupture force, rupture displacement, yield work, yield force and yield displacement.Factor analysis was applied to these indicators,and the original indicators were reduced in dimensionality,so that a comprehensive index was used to evaluate 1-MCP(1.0 μL/L)treatment,it was placed at shelf temperatures of 5,10,15,20 ℃ to maintain the texture properties of Huahong and Huayue apples.The shelf life of Huahong and Huayue fruit texture characters could be prolonged by 1-MCP treatment,especially for Huahong fruit.Under the same treatment temperature,the fruit decline of the two varieties treated with 1-MCP was significantly less than that of the untreated group,which could be reduced by about 10%.Correlation analysis showed that there was a very significant positive correlation between the texture indexes of apple fruit,but there were some differences in the degree of tightness.After factor analysis and based on the principle that the eigenvalue was greater than 1,two main factors were extracted,and the cumulative variance contribution rate was 91.194%.According to the representative indexes of each main factor,they were named puncture factor F₁ and TPA factor F₂,and the variance contribution rates were 79.870% and 11.324%,respectively.According to the score of the main factor,the most suitable shelf temperature of untreated Huahong fruit was 5 ℃.The temperature of the treated fruit was 15 ℃.The optimum shelf temperature of Huayue fruit without treatment was 10 ℃,and the suitable shelf temperature of 1-MCP treated fruit was 5,10 ℃.The CK group of Huahong fruit could be stored for 8 d at 5 ℃,and the shelf temperature of the treatment group could be extended to 16 d at 15 ℃;the CK group and the treatment group of Huayue fruit could be stored at 10 ℃,could be stored for 16 d.

The aim of this study was to investigate effects of wine quality in wine grapes Marselan on different rootstocks and provided theoretical reference for cultivation regulation.Wine grape Marselan grafted was used as experimental material in 2018,rootstocks were SO4,5BB,3309M and 101-14,respectively,own-rooted grape was used the control.The fruit was harvested when ripe in 2020 and 2021,bunch character and wine quality were measured.The results showed that density, bunch weight, average berry weight, berry length and berry width, maximum berry weight and No. of shriveling berry decreased,however,length and width and average berry weight of CK increased.Length and width,average berry weight and average seed number increased by grafting,the coefficient of variation of berry quality was small in 2 years.The berry weight of M/SO4,M/5BB and M/3309M mainly ranged from 1.01 to 1.25 g,M/101-14 mainly ranged from 0.76 to 1.00 g in 2020.The berry weight of M/5BB,M/3309M and M/101-14 mainly ranged from 0.76 to 1.00 g,M/SO4 mainly ranged from 1.01 to 1.25 g in 2021.Soluble solids decreased,however total acid mass concentration increased by grafting.Component and content of anthocyanins in peel were measured by LCMS/MS,the results showed that malvidins content was the highest,the second was delphinidins,pelargonidin was the least.Anthocyanins contents of M/5BB significantly increased by 26.62% compared with CK,anthocyanins contents of M/3309M and M/101-14 increased,but there was no difference with CK,anthocyanins contents of M/SO4 significantly decreased by 11.87% compared with CK.

In order to study the application method of humic acid on grapes and find out the optimal proportion of carbon such as humic acid instead of organic fertilizer,the plot experiment was carried out with simple chemical fertilizer fertilization mode as control(CK treatment)for two consecutive years.Based on farmers' fertilizer practice(FFP treatment),3 ratios of carbon such as humic acid instead of organic fertilizer were set:10% carbon substitution(T1 treatment),20% carbon substitution(T2 treatment)and 30% carbon substitution(T3 treatment).Their effects on grape yield,quality and soil nutrients were studied,and the optimal substitution ratio was screened by principal factor analysis.The results showed that the yield of T2 treatment was the highest for two consecutive years,but there was no significant difference between T2 and T3 treatment.Single cluster weight and single fruit weight in T1,T2,T3 treatments were significantly higher than those in CK treatment,but there was no significant difference between FFP treatment and T1,T2,T3 treatments;on the whole,the contents of soluble solids,soluble sugar,titratable acid and vitamin C in humic acid treatment instead of organic fertilizer were better than those in other treatments.Through correlation analysis,soil available potassium,organic matter and acid phosphatase were positively correlated with grape yield.Available phosphorus,available potassium,organic matter and acid phosphatase in soil were very significantly correlated with soluble solids,soluble sugar and vitamin C content of grape.Regulation of these soil indexes was the way to increase grape yield and quality by replacing organic fertilizer with humic acid;through principal factor analysis,there were 2 principal factors with eigenvalue greater than 1 in 2019 and 2020,the cumulative contribution rate of variance of 2 principal factors reached 88.864%-91.470%,and T2 treatment was the highest comprehensive score for two consecutive years.On the whole,carbon such as humic acid instead of 20% organic fertilizer was the optimal proportion.

Gene STAYGREEN(SGR)plays an important role in the process of plant chlorophyll degradation and metabolism.To explore the biological function of gene SGR2,MdSGR2 gene sequence was cloned from the peel of Granny Smith by RT-PCR.The homology,physicochemical properties,protein structure and cis-acting elements of the promoter were predicted and analyzed by bioinformatics software.The plant overexpression vector was constructed by double enzyme digestion.The results showed that the complete open reading frame(ORF)cDNA of gene MdSGR2 was 840 bp.Encoding 279 amino acids in total,it belonged to the Staygreen superfamily.Physicochemical analysis showed that the total molecular weight of the protein was 31.27 ku,and the isoelectric point pI was 8.52,indicating that the protein was an unstable hydrophilic protein.Homology analysis showed that MdSGR2 encoded amino acid sequence was of the highest homology with Pyrus ussuriensis,reaching 84.12%.The protein structure analysis showed that the secondary structure and tertiary structure of MdSGR2 was determined structurally to be 40.55% α-helix and 44.44% random coil.Promoter analysis showed that the promoter region of the gene contained cryogenic cis-acting elements,photo-responsive elements and induced cis-acting elements such as abscisic acid,salicylic acid and methyl jasmonate response elements.These results suggest that gene MdSGR2 may be regulated by many factors such as environment and hormones.The plant overexpression vector pCAMBIA2301-MdSGR2 was successfully constructed.

In order to reveal the effect and molecular mechanism of salicylic acid-induced resistance to black spot disease of Yali,the leaves and fruits of Yali were used as experimental material which were collected from the experimental orchard of Hebei Agricultural University.Tissue isolation method was used to isolate and purify the pathogen of black spot disease and detected its pathogenicity.Morphological observation and multi-genes of ITS,HIS,RPB2 and ACT were used to identify the pathogen of black spot disease.The detached leaves of Yali were inoculated with conidia suspension of Alternaria alternata by acupuncture for 0,6,12,24,48,72,96 and 120 h,respectively.Real-time RT-PCR was used to analyse the expression of related genes which involved in salicylic acid signaling,and high-performance liquid chromatography (HPLC) was used to detect the content of endogenous SA at 0 and 72 h,respectively.Exogenous SA treatment with different concentrations (0,0.002,0.02,0.2,2.0,10.0,20.0 mmol/L) were used to identify the effect to the mycelial growth of A.alternata;Exogenous SA treated Yali fruit with different concentrations (0,0.02,0.2,2.0 mmol/L),respectively,and inoculated with A.alternata to detect the effect of disease resistance.The results showed that the pathogen of Yali black spot disease was A.alternata. Detached leaves of Yali were inoculated with A.alternata for 72 h,the free state SA content increased from 0 mg/g to 0.02 mg/g and the binding state SA content increased from 0.47 mg/g to 1.55 mg/g,and for 96 h compared with 0 h,the expression of Pbrgene12425,Pbrgene6286, increased 5.48,4.66 times,respectively,the expression of Pbrgene8895 and Pbrgene43605 increased 7.90 and 10.0 times for 120 h.Exogenous 0.2 mmol/L SA significantly increased the resistance of Yali fruit to black spot disease.In the process of Yali resistance to black spot disease,the free state SA content of Yali leaves significantly increased,and the related genes in SA signaling were induced to up-expressed,exogenous 0.2 mmol/L SA significantly increased the resistance of Yali to black spot disease.

The suitable means to construct core collection of Yanshan chestnut germplasm based on SSR makers was studied, which was beneficial for preservation, management and utilization of chestnut germplasm resources.Taking 161 Yanshan chestnut germplasm resources from 10 cities(counties)as original materials, an allele-preferred sampling method and random sampling strategy were compared using UPGMA cluster method according to 3 genetic similarity coefficients(SM, Dice and Jaccard coefficient)based on SSR molecular markers. The effective number of alleles(Ne), Nei's gene diversity(H)and Shannon's information index(I)of two groups were compared to determine the optimum method. t -tests of core collection, initial collection and reserve collection were conducted to evaluate the representativeness of core collection. The principle coordinate analysis method and phonotypic traits analysis were used to confirm on the core collection.Compared with the random sampling strategy, allele preferred sampling method was more representative with higher values of genetic diversity indexes. The results showed the method of stepwise clustering according to SM similarity coefficient was better than Dice coefficient and Jaccard coefficient. 46 core collection included 28.57% initial collection. The effective number of alleles, Nei's gene diversity, Shannon's information index were respectively 1.531 7, 0.321 8 and 0.491 0. t -test showed the genetic diversity index of core collection was significantly higher that of initial collection.The core collection was evenly distributed in the principle coordinate diagram of initial collection, which could represent genetic diversity of the whole chestnut germplasm comprehensively.The method of allele preferred sampling and stepwise clustering according to SM similarity coefficient was a suitable means to construct core collection of Yanshan chestnut germplasm based on SSR makers, and 46 core collection could represent genetic diversity of original germplasm resources.

The study on the diversity and variation of fruit phenotypic characters of walnut germplasm resources can provide valuable based data and theory foundation for construction of walnut germplasm resources and variety breeding, and promote the efficient utilization of walnut germplasm resources. The study was conducted in order to evaluate 13 qualitative characters and 11 quantitative characters on 99 walnut germplasm resources, which were designed with hierarchical classification and then were done with analysis of variations and systematic cluster analysis. The results showed that the coefficient of variation of the 13 qualitative characters ranged from 16.57% to 54.46%, especially, coefficient of variation concerning 7 out of 13 characters including sweet, texture of crisp, number of kernel, number of pockmark, degree of difficulty taking kernel, astringency and smooth of shell. The Shannon-weaver index of the 13 qualitative characters ranged from 0.49 to 1.51, especially, Shannon-weaver index concerning 5 out of 13 characters including degree of difficulty taking kernel, number of kernel, smooth of shell, color of shell and astringency. The coefficient of variation of the 11 quantitative characters ranged from 8.38% to 26.91%, especially, coefficient of variation concerning 2 out of 11 characters including weight of kernel and thickness of shell. The Shannon-weaver index of the 11 quantitative characters ranged from 1.89 to 2.27, especially, Shannon-weaver index concerning 3 out of 11 characters including equatorial diameter of nut(2.27), side diameter of nut(2.27), thickness of shell(2.27). There were significant differences among and within populations(except nut weight and equatorial diameter of fruit), and phenotypic differentiation coefficient among and within populations were 15.66% and 58.13% respectively. The degree of difficulty taking kernel had extremely significant correlation with thickness of shell(r=-0.618), kernel rate(r=0.573), number of kernel(r=0.790), degeneracy of crisp(r=0.733), texture of crisp(r=0.659). The systematic cluster analysis showed that 99 test materials could be divided into 3 categories based on the variation of phenotypic characters and in which there were differences. There were abundant genetic diversity based on the phenotypic characters of fruit of walnut, and these characters were worth mentioning such as degree of difficulty taking kernel, number of kernel, smooth of shell, astringency, thickness of shell, degeneracy and texture of crisp in the variety breeding. The variation within populations was more abundant(except the fruit shape index), and we can pay attention to the genetic improvement within populations.

The changes of minerals in leaves(L), flowers/fruits(F)and their correlations in the full-bloom stage(FBS), veraison stage(VS), and maturation stages(MS)of Merlot grapes in the Penglai producing area was analyzed, and which will provide the theoretical basis for the nutritional diagnosis and scientific fertilization of Merlot grapes. In 2018-2020, 16 Merlot grapes under fixed formula fertilization in the Penglai production area were selected as the research object. Each formula fertilizer selected 4 orchards, a total of 192 Merlot vineyards, and the leaves, flowers/fruits of different growth periods were compared. For differences in the content of mineral elements, correlation analysis was used to screen the main leaf nutrient factors that affect the mineral nutrient content of the fruit. The contents of L_Fe, B and F_N, P, K, Mg, Fe, and B content in each growth period showed the order of FBS>VS>MS, and there were significant differences in different growth periods. The contents of L_Mn, Cu and Cu content showed the order of VS>MS>FBS, and the differences were significant in each growth period. Except for N/Ca/Mg at FBS, Cu at MS, and Mn at FBS NS/MS, other leaf and flower/fruit mineral elements showed significant or extremely significant differences. The contents of N, Ca, and Mo in each growth period showed the order of L>F. The content of Fe, Mn, Zn, and B in each growth period showed as F>L. The content of P and K was expressed as F>L in both FBS and VS, and L>F in MS. The content of Mg and Cu in FBS was F>L, and in VS and MS, the content was L>F. There were similarities and specificities in the changes of mineral elements in the leaves and fruits of Merlot grapes, which might be caused by the different degrees of synergy or antagonism between the various mineral elements in the leaves and fruits. Correlation analysis showed that the content of various elements in leaves and fruits showed different degrees of correlation during different growth periods. P, K, Fe, Zn, Cu, B were indispensable elements in the development of Merlot grape flowers. The content of Ca, Mn, and Cu in the leaves and fruits of Merlot grapes increased significantly during the period of FBS and VS. During the MS of Merlot grapes, the N, P, K, Ca, Mg, and Mo contents in the leaves were significantly higher than those in the fruits, and the contents of Ca, N, and K were higher, indicating that the effects of Ca, N, K in the growth and development of the fruit were relatively high. The demand was higher than other mineral elements. In summary, the P, K, Ca, Mg, Fe, Zn, Cu, B, and Mo levels in the leaves before anthesis could be adjusted to regulate the content of P, K, Fe, Cu, and B in the flower;because the Ca in the leaves was not easy to effectively transport to the fruit for its use, so Ca should be directly applied to the young fruit in a targeted manner. During the VS, the content of Mn and Cu in fruits could be regulated by increasing the levels of Mn and Cu in leaves. During maturity, the contents of P, K, Zn, and Mn in the fruit could be regulated by increasing the levels of N, K, and Ca in leaves.

The effects of 1-MCP and ethephon on the main physiological quality indexes and softening related indexes of Postharvest Pyrus ussuriensis during normal temperature storage were studied, and the mechanism of 1-MCP and ethephon on softening regulation of Pyrus ussuriensis was discussed, so as to provide theoretical basis for regulating the ripening and softening process of Pyrus ussuriensis. Five kinds of Pyrus ussuriensis, Nanguo pear, Jingbai pear, Huagai pear, Jianba pear and An pear, were fumigated with 0.5 μL/L 1-MCP for 24 h and 1.0 g/kg ethephon solution respectively. The changes of fruit firmness, respiration rate, ethylene production, soluble solids, titratable acid, vitamin C content, water-soluble pectin, cellulose, starch content and softening related enzyme activities of polygalacturonase(PG), β-galactosidase(β-Gal), cellulase(CL)and amylase(AM)were studied during normal temperature storage((20±1)℃). 1-MCP treatment significantly delayed the decline of firmness, ethylene production, respiration rate, water soluble pectin(WSP), cellulose and starch content of Pyrus ussuriensis, and decreased the activities of PG, β-Gal, CL and AM enzyme, but there was little difference between ethephon treatment group and control group. 1-MCP treatment could alleviate the degradation of cell wall substances by inhibiting the activities of cell wall related degrading enzymes and reducing ethylene production, thus effectively delaying the fruit softening process of Pyrus ussuriensis. However, ethephon had no significant effect on the physiological changes of fruit softening and cell wall degradation during storage.

To explore the effects of spraying CaCl₂ at different concentrations on physiological characteristics of apple seedlings under salt stress, two-year-old T337 seedlings were used as test materials, and the pot experiment was used to set 100 mmol/L NaCl(T1) and 100 mmol/L NaCl + four different concentrations of CaCl₂ (5.0(T2), 7.5(T3), 10.0(T4), 12.5 mmol/L(T5)) treatment, watering with deionized water was used as the control(CK).Determine the chlorophyll content, photosynthetic and fluorescence parameters, antioxidant enzyme activity and osmotic adjustment substance content of each treatment group, and carry out correlation and principal component analysis. The results showed that with prolonged stress time, the catalase(CAT), peroxidase(POD), superoxide dismutase(SOD) of T337 leaves first increased and then decreased, malondialdehyde(MDA), intercellular CO₂ concentration(Ci), proline(Pro) and relative conductivity(REC) showed a rising trend, initial fluorescence(F0), PSⅡ maximum light energy conversion rate(Fv/Fm) stomatal conductance(Gs) and the transpiration rate(Tr) showed a downward trend;compared with CK and T1, the variation amplitudes of T337 leaves after treatment with exogenous CaCl₂ were all reduced, and there was a significant concentration effect. Among them, the relative chlorophyll content(SPAD), Pn, Tr, POD, F0 and SOD decreased the least under T4(10mmol/L) treatment, which was significantly higher than other treatment groups;MDA, REC and Ci increased the least, which was significantly higher than CK and lower than others processing group. Correlation analysis of 14 indicators showed that Pn was positively correlated with CAT, Tr, Gs, F0, Fv/Fm, ETR, SPAD, SOD and POD, and negatively correlated with Pro, MDA, Ci, and REC. Principal component analysis was performed on 14 related indicators after salt stress treatment, and 2 principal components with eigenvalues greater than 1 were extracted, and their eigenvalues were 11.190, 2.295, respectively;The first and second principal component variance contribution rates were 79.930%, 16.394%, respectively, the cumulative variance contribution rate reached 96.324%, which meets the analysis requirements. According to the ranking of principal component scores, the mitigation ability of exogenous CaCl₂ to T337 salt stress was T4>T3>T2>T5 from high to low. Therefore, 10 mmol/L CaCl₂ could improve the photosynthetic fluorescence characteristics of T337 rootstock leaves under salt stress, improve the antioxidant enzyme system and osmotic adjustment ability, and enhance the stability and permeability of biofilms to alleviate salt stress.

In order to investigate the effects of different materials of modified atmosphere packaging on the postharvest physiology and storage quality of Hongxiangsu pear, six kinds of packaging treatments, such as CK, high exudation CO₂ bag, PE bag, PE bag + 1-MCP preservative, PE bag + ethylene absorbent, PVC bag, were studied. The fruits of Hongxiangsu pear were refrigerated at (0±0.5)℃ for 120, 180, 240 d. After being taken out, the fruits were balanced at 20℃ for 24 h and stored for 7 d. The changes of respiration rate, ethylene release, ethanol and acetaldehyde contents, fruit appearance and internal quality were measured and the contents of CO₂, O₂ and ethylene in different bags were monitored regularly. The results showed that compared with CK and high exudation CO₂ bag, PE bag and PVC bag had good air conditioning ability, which could significantly reduce the volume fraction of O₂ and increase the volume fraction of CO₂, but the ethylene concentration in the bag was also relatively high, while the ethylene concentration in PE bag + ethylene absorbent bag was relatively low. Compared with CK fruit, the other four kinds of packaging treatments except high exudation CO₂ bag could delay the decline of fruit firmness and the yellowing of peel in the middle and later storage periods, maintain relatively high soluble solid content. Among them, the PE bag+1-MCP preservative not only maintained better appearance and normal taste flavor of Hongxiangsu pear fruit during cold storage and shelf life, but also significantly reduced the fruit respiration rate, ethylene release, mass fraction of ethanol and acetaldehyde. However, when the fruits were stored in PVC bags for 240 d and stored at 20℃ for 7 d shelves, some fruits showed browning and abnormal flavor. For Hongxiangsu pear stored in medium and long term, the PE bag+1-MCP preservative treatment had the best storage effect, followed by single PE bag and PE bag + ethylene absorbent. PVC bag was not recommended for Hongxiangsu pear.

To understand the effects of SH1 and Y-1 dwarfing interstocks on the growth and leaf photosynthesis of Fuji apple saplings, and provide theoretical basis for the selection and utilization of dwarfing interstocks in production, the Fuji Nagafu 2 apple trees grafted on SH1 and Y-1 interstocks were used to study the effects of two kinds of interstocks on the growth, branch composition and leaf photosynthetic indexes of young Fuji trees. The results showed that the annual growth trend of Fuji on SH1 and Y-1 interstocks was basically the same, but the growth of spring and autumn shoots and the length of new shoots of Y-1 were significantly lower than that of SH1, and the ratio of short branches was significantly higher than that of SH1, and the fruit color index, hardness, soluble solids and solid acid ratio significantly increased. The diurnal and annual dynamic trends of net photosynthetic rate(Pn) of Fuji Nagafu 2 were different between SH1 and Y-1 interstocks. The Pn of leaf in SH1 in the early growth period was higher than in Y-1, but the Pn of leaf in Y-1 in the later growth period was significantly higher than in SH1, analysis of light response parameters and chlorophyll fluorescence parameters showed that the light saturation point(LSP) of Y-1 in the early growth period was lower than SH1, the compensation point(LCP) was higher than SH1, non photochemical quenching coefficient (NPQ) was significantly higher than SH1, and the primary light energy conversion efficiency (Fv/Fm), photosynthetic electron transfer quantum efficiency (ΦPSⅡ) and apparent electron transfer rate (ETR) of Y-1 were significantly lower than that of SH1 in the early growth stage, and the LSP, PSⅡ, Etr, the chlorophyll content and net photosynthetic rate of Y-1 were significantly higher than those of SH1 at the later growth stage. Different intertstocks had great effects on the growth and leaf photosynthetic characteristics of grafted varieties. Some characteristics of Fuji Nagafu 2 grafted onto Y-1 were better than that of SH1, such as dwarfing, early flowering, easy shaping. In the early growth period, the leaf maturity of Fuji Nagafu 2 in Y-1 lags behind in SH1, so the photosynthetic efficiency of the leaves was low. In the late growth period, the photosynthetic pigment accumulation and Fv/Fm of leaves in Y-1 were higher or significantly higher than that of SH1, and it could effectively control the growth of autumn shoots, which was beneficial to the accumulation of nutrients and the formation of flower buds.

In order to provide reference for the selection of suitable scion-rootstock combinations in the production of wine grape in Ningxia.Chardonnay and Merlot were grafted with three rootstocks respectively to study the effects of rootstocks on the growth, fruit quality and yield of two kinds of grapes.The results showed that 5BB, 1103P and 110R promoted the growth of Chardonnay in different degrees, among which 1103P increased the trunk diameter, leaf area and SPAD value significantly, and the effect of increasing growth was the most obvious. 5BB significantly increased the reducing sugar and titratable acid content of Chardonnay fruit, while 1103P decreased the accumulation of reducing sugar and increased the content of titratable acid, and 110R had no significant effect on the sugar and acid content. 1103P and 110R significantly reduced the total phenol content of Chardonnay, while 5BB and 1103P increased the anthocyanin content significantly. There was no significant difference in yield per plant between the grafted and self-rooted Chardonnay. The growth of Merlot grafted onto 140R, 5BB and SO4 was slowed down evidently, among which 140R significantly reduced the trunk diameter, new shoot length and diameter, leaf area and SPAD value, and the effect of weakening growth was the most obvious. SO4 significantly decreased the reducing sugar content and increased titratable acid content of Merlot fruit, while 5BB had no significant effect on the reducing sugar content, but significantly increased the titratable acid content, and 140R had no significant effect on sugar and acid content. Three rootstocks improved the tannin and anthocyanin content of Merlot fruit, among which 140R showed a significant raise in tannin content, and the anthocyanin content of SO4 and 140R grafted was obviously higher than that of self-rooted. The rootstock 5BB significantly increased the yield per plant of Merlot.Among the three rootstocks, 5BB was the best one for Chardonnay;but for Merlot, the self-rooted seedling was the best, and the comprehensive performance of 5BB was better than the other two rootstocks in terms of stable growth, quality and yield.

In order to explore the relationship between the superficial scald and related physiological indexes of Dangshansu pear during storage, the mineral elements(N, P, K, Mg, Ca) content, respiration intensity, ethanol content, relative conductivity(cell membrane permeability), total phenol content, polyphenol oxidase activity(PPO activity), chlorophyll fluorescence parameters and related physiological indexes of Dangshansu pear fruits with different levels of superficial scald were measured, and the correlation between the various physiological indicators and the superficial scald progression was analyzed. The results showed that the content of Ca in fruits of different grades of superficial scald was significantly lower than that of normal fruits(no superficial scald), while the content of N, K, Mg and P were higher than that of normal fruits. There was an extremely significant positive correlation between the incidence degree of superficial scald and N content, and an extremely significant negative correlation between the incidence degree of superficial scald and Ca content. Results showed that the fruit respiration intensity, ethanol content(peel), relative electric conductivity(flesh), total phenol content(peel and core), PPO activity(peel, flesh, core) and incidence degree of superficial scald were very significant positive correlation, both the chlorophyll fluorescence parameters(Fo, Fm, Fv, Fv/Fm, Fv/Fo) of normal fruit peel were significantly higher than superficial scald fruit. It was preliminarily determined that the higher N content and the lower Ca content in the fruit were the main causes of the superficial scald in the late stage of storage. The more serious the superficial scald was, the higher the respiration intensity, ethanol content, tissue membrane permeability and PPO activity were, and the lower the chlorophyll fluorescence parameters of peel(Fm, Fo) were. Therefore, the occurrence of superficial scald during storage can be reduced by regulating mineral nutrients or post harvest calcium supplementation before harvest, so as to improve fruit storage quality.

The browning of sweet cherry fruit not only affects its nutritional value but also affects its commerciality.In order to study the relationship of browning and the expression level of PPO,POD,CAT,SOD genes under different EBR concentration treatment,sweet cherry were used as test materials.Changes of fruit browning index,PPO,POD,CAT,SOD activity and relative expression of PPO,POD,CAT and SOD genes were measured during storage.The results showed that the fruit browning index of the control group was significantly higher than that of the EBR treatment groups after 30 days of storage,and the fruit browning of the control group was more serious.The activity of PPO and POD in the control group were always higher than that in the EBR groups during the whole storage process.CAT and SOD activity in EBR treated fruits were always higher than that in untreated fruits after storage for 15 days.The relative expressions of PPO and POD genes in the control group were significantly higher than those in the EBR treatment groups.The relative expressions of CAT and SOD genes in EBR treated fruits were significantly higher than those in untreated fruits after storage for 30 days.The above results showed that EBR treatment could inhibit the relative expression of PPO and POD genes and reduce the activity of PPO and POD.It could promote the relative expression of CAT and SOD genes and improved the activity of CAT and SOD.The effect of 5 μmol/L EBR treatment was better,which could maintain the quality and flavor of sweet cherry fruit.

In order to illuminate the mechanisms of fruit cracking of Chinese jujube under water stress,5 a Jinsi novel No.4 Chinese jujube was chosen.The trees were spayed for 50 hours to simulate water stress.Fruits before and after treatment were sequenced on the ONT third-generation sequencing platform.There were totally 13 368 transcripts obtained from 6 samples.9 178 PolyA sites were identified in the APA event.78 significantly enriched PolyA signals(PAS)motif were found.7 321 SSRs and 114 lncRNAs were predicted.239,189 alternative splicing events were identified in the control and treatment group,respectively.There were 248 differentially expressed transcripts(DETs)between the two groups.DETs mainly encoded calnexin,calreticulin,heatshock proteins, microtube proteins fructokinase and so on.GO enrichment showed that DETs were mainly involved in the biological processes such as translation,positive regulation of mitotic centrosome separation and cold acclimation.Cellular components participating those processes included cytosol and cytosolic large ribosomal subunit etc.Enriched molecular functions included structural constituent of ribosome and xyloglucosyl transferase activity,etc.KEGG enrichment analysis showed that up-regulated DETs were involved in protein processing in endoplasmic reticulum,phagosome,amino sugar and nucleotide sugar metoboism,starch and sucrose metabolism,etc.Down-regulated DETs took part in protein processing in endoplasmic reticulum,nitrogen metabolism,photosynthesis-antenna proteins and so on.The mechanisms of fruit cracking of Chinese jujube under water stress might include decrease of alternative splicing events,regulation of Ca²⁺ by calnexin and calreticulin,up-regulation of heat shock proteins resulting from cold stress by continuous spaying,and up-regulation of cell wall biosynthesis related transcripts.

Genetic variations analysis of 127 Yanshan chestnut germplasms from 11 populations 15 phenotypic traits related catkin of Yanshan chestnut were designed with genetic diversity analysis,correlation analysis and principal component analysis,in order to provide valuable basic data and theory foundation for normalization,standardization,preservation and construction of chestnut core collections,provides the basis for the gene localization of agronomic traits. The results showed that:phenotypic traits related catkin of Yanshan chestnut were rich in phenotypic diversity,with mean variation coefficient (CV) of 27.55% and Shannon's diversity index(H')of 1.99. The mean variation coefficient(CV=64.80)of bisexual catkin numbers/catkin numbers was the largest,the mean variation coefficient (CV=7.52)of male catkin numbers/catkin numbers was the smallest. The Shannon's diversity index(H'=2.08)of stamen length was the highest,the Shannon's diversity index(H'=1.89) of bisexual catkin numbers/catkin numbers were the lowest. 9 phenotypic traits related to catkin were highly significant which indicated that a wide range of variation existed in these two levels. Correlation analysis showed that:More flower traits were highly significantly and extreme significantly related. Principal component analysis showed that floral traits in the first 5 principal components reflect the total information content 85.35%,bisexual catkin numbers/bearing shoot,catkin length/catkin width,catkin width,catkin length,bisexual catkin numbers/catkin numbers,male catkin numbers/catkin numbers,female flower numbers/bearing shoot,stamen length,floral axis length/floral axis width,9 characters was the main factor causing variation in traits of Yanshan chestnut. The 127 materials were divided into 8 kinds of cluster analysis. Flower character same source with similar material part can get together,the results were similar to principal component analysis.

Cold stress is a major environmental factor that limits peach growth,development reproduction and distribution. Clarifying the molecular mechanism of peach in response to low temperature stress could provide the research basis for cultivating and screening relatively cold resistant peach varieties. In cold-tolerant peach cultivar Qiuyan were selected as testing materials. 1 year dormant branches were exposed to-5(the control),-15,-25℃under controlled environment chamber for 48 h respectively. Two-dimensional electrophoresis(2-DE)were used to analyze proteomics of cortex of branches under 3 different low temperature treatments. The results showed that a total of 150 differentially expressed proteins were identified by mass spectrometry,97 significantly up-regulated proteins and 53 significantly down-regulated after cold stress. After further analysis of the protein spots by proteome data retrieval,20 proteins related to cold stress were identified,involving amino sugar and nucleotide sugar metabolism,starch and sucrose metabolism,amino acid metabolism,MAPK signaling pathway,phenylalanine metabolism and DNA replication. In addition,Real-time PCR (RT-PCR) indicated that three candidate protein-encoding genes Alpha-amylase(M5VVU6),Chlorophyll a/b binding protein(M5WUQ0)and Epoxide hydrolase 3(D8L7V9)were up-regulated with the increase of cold stress. It suggested that these metabolic pathways and three specific proteins might play an important role in the cold-resistant mechanism in peach.

In order to explore the effect of grafting on the volatile flavor compounds of yellow cherry tomato fruit,the volatile flavor compounds and relative contents of three yellow cherry tomato varieties were analyzed by headspace solid phase micro-extraction with gas chromatography-mass spectrometry. The results showed that there were 56 volatile compounds in Huangrong S,including 3 unique volatile compounds; there were 56 volatile compounds in Huangrong J,including 3 unique volatile compounds. Jinlulu S contained 46 volatile compounds,including 4 unique volatile compounds,of which 1-nitro-3-methylbutane was the main flavor compound; Jinlulu J contained 57 volatile compounds,including 15 unique volatile compounds,of which heptanal,benzeneacetonitrile,1-nitro-2-phenylethane and β-ionone were the main flavor compounds. Huangxing S contained 58 volatile compounds,including 7 unique volatile compounds,of which benzeneacetaldehyde,benzeneacetonitrile and 1-nitro-2-phenylethane were the main flavor compounds; Huangxing J contained 61 volatile compounds,including 10 unique volatile compounds,of which guaiacol was the main flavor compound. It was found that grafting significantly affected the categories and relative contents of volatile compounds in the three yellow cherry tomato varieties fruit. Compared with the non-grafted plant,the relative content of 2-methyl-1-butanol decreased while the relative content of β-ionone,E-2-heptenal and Z-4-decenal increased in the grafted fruits. This study laid a foundation for further analysis of tomato fruit flavor metabolism.

For the problem of soil desertification caused by strong wind for long time and development of local grape industry in Yanhuai Basin,the influence of grasses on soil physical and chemical properties in the vineyards of Yanhuai Basin was determined,and the technology of planting grasses in vineyard was explored. Two kinds of grasses were selected for experiment and planted in 2017 according to the climatic conditions and characteristics of viticulture in Huailai County,and traditional cultivation as control. The physicochemical properties of sandy soil were analyzed after 3 years of planting. The results showed that:Compared with control,the nitrate content of 0-20 cm surface soil was increased significantly,and the available phosphorus,total nitrogen content of 0-80 cm soil layer was increased after planting Orychophragmus Violaceus (L.)O.E. Schulz for 3 years,while the physical properties of surface soil was improved.Compared with control,the organic matter and total nitrogen content of 0-20 cm surface soil was increased significantly after planting Carex lanceolata Boott. After 3 years of continuous cultivation,the contents of carbon and nitrogen in the aggregate of the surface soil were increased in vineyard.The influence of the two kinds of grasses on the aggregate structure of the sandy soil of the vineyard was different. The macroaggregate(>0.25 mm)content of surface soil was increased significantly after planting Carex lanceolata Boott and the microaggregate(<0.25 mm)content of surface soil was increased significantly after planting Orychophragmus Violaceus (L.)O.E. Schulz,so it was suggested that the surface soil should not be artificially ploughed after planting Orychophragmus Violaceus (L.)O.E. Schulz in order to prevent soil erosion in Yanhuai Basine. Soil physical and chemical properties could be improved and the effects on soil physical and chemical properties was different after planting grasses in vineyard,but the selection of suitable grass species was important in vineyard of Yanhuai Basin.

To explore the botanical agents to prevent and control apple ring spot disease,the inhibitory of the four common of Allium plant extracts,on the hypha growth of Botryosphaeria berengriana were tested.On this basis,the metabolites of the four Allium plants,Chinese leek,garlic,onions and welsh onion,were analyzed by metabolomics methods,and the suppressive effects of the main components of the four Allium plants on the mycelial growth of B.berengriana and the incidence of the apple ring disease on the apple fruits were also determined.The results showed that in the whole experiment period the inhibition of four Allium plant extracts(1×)on the growth of mycelia of B.berengriana was as high as 100%.78 metabolites were detected in the four Allium plants.Among them,the three main components,pyroglutamic acid,phosphoric acid and citric acid,inhibted the growth of mycelia of B.berengriana by 29.10% to 100.00%,and inhibited the incidence of ring disease on apple fruit caused by B.berengriana by 44.38% to 91.45%.The results of this study laid a theoretical foundation for further understanding and developing the antimicrobial components in Allium,and provided a potential way for the prevention and treatment of apple ring disease.

This study aims to explore the genetic diversity of Malus at the level of chloroplast gene and the phylogenetic relationship of haplotypes of different species in different regions, so as to provide a basis for its origin and evolution process, protection and utilization. Based on chloroplast intergenic regions trnH-psbA, trnS-trnG spacer+intron, trnT-5'trnL and 5'trnL-trnF, the sequences of 722 accessions of Malus from twelve provinces were analyzed. After four regions merged, the length of four merge regions of chloroplast DNA was 4 120 bp, and the haplotypes of chloroplast DNA fragments were 100, the nucleotide diversity and haplotype diversity of the merged regions were 0.009 52 and 0.879 respectively. The region with highest nucleotide diversity and haplotype diversity was trnH-psbA (Hd=0.808,Pi=0.034 09). Tajima's test showed that the value of four regions merged was -1.503 16, and it was not significant at the P>0.10 level and followed the neutral theory of molecular evolution. AMOVA showed that genetic variation mainly existed among populations and within intraspecific populations.The results of haplotype distribution and network analysis showed that the missing haplotypes in the center of adjacent networks are closed into a ring, and different species of Malus have different evolutionary routes. There were complex relationships among species and germplasms of same species in the process of origin and evolution in different regions. Relatively old haplotypes H_6 and H_15 experienced population expansion, and there were many derived branches. The genetic diversity of Malus at chloroplast gene level was high, and the genetic variation mainly occurs among populations and within intraspecific populations. The genetic evolution of chloroplast genes was dominated by mutation or random drift within populations. The main factors of genetic variation of Malus were intraspecific differences and intrapopulation differences caused by geographical isolation.

For the development of a large number of SNP for the construction of high quality genetic map of cultivated strawberry(Fragaria×ananassa,2n=8X=56),Hongxing×Benihoppe crossing populations were sequenced by SLAF-seq that yielded 565 919 066 reads, with Q30 value of 95.36% and GC content of 39.68%. 717 881 SLAFs and 2 136 939 SNPs were identified.Total 56 237 SNPs showed high quality for genetic map construction, of which 14 412 were successfully mapped to 28 linkage groups spanning a total of 4 022.16 cM with an average interval genetic distance of 0.28 cM. It was showed that the greater part of each individual was derived from the parents by evaluating the monomer source of the genetic map. The results of heat map showed that the sequence of most markers on each linkage group was consistent with the genome, the sequence of markers was correct, which could show that the map was of high quality.A high-density genetic map of cultivated strawberry by SLAF-seq could be laying a foundation for genetic research and molecular marker-assisted breeding of cultivated strawberry.

In order to study the effects of nitrogen application rate on the photosynthetic characteristics and assimilation product accumulation of Cabernet gernischet grapes at different growth stages, four urea application rates under drip irrigation conditions were set:150(N1), 300(N2), 450 (N3), 600 kg/ha(N4), 0 kg/ha was set to control(CK), the effects of different nitrogen nutrition on photosynthetic fluorescence parameters and carbon metabolism related indexes of grape leaves at growth period (5 days before flowering,20 days after flowering,50 days after flowering) were analyzed. The results showed that appropriate nitrogen application improved the chlorophyll content and RuBp carboxylase activity of leaves, enhanced the photosynthesis of vines, and increased the chlorophyll fluorescence of leaves which represented as the increase of effective photochemical efficiency (Fv'/Fm'), photochemical quenching coefficient (qP) and actual photochemical efficiency of PSⅡ(ΦPSⅡ) and the decrease of non-photochemical quenching coefficient (NPQ) in leaves. Compared with CK, Fv'/Fm', ΦPSⅡ, and qP under N2 treatment in DAF20 and DAF50 were significantly increased by 16.10%, 17.80%, and 21.49%, 21.13%, and 11.77%, 19.73%, respectively.Appropriate nitrogen application increased the activity of sucrose metabolism-related enzymes and soluble sugar content, but decreased starch content.And throughout the growth period, SPS, SS, and sucrose had the highest values under N3, glucose and fructose had the highest values under N2, while NI and AI had the highest values under N2, N2 and N1 respectively under DBF5, DAF20 and DAF50. Compared with CK, N2 treatment increased the content of soluble sugar, tannin, anthocyanin and the titratable acid content, promoted the fruit setting rate and the ratio of sugar to acid with yield of 14.50 t/ha.In conclusion, the application range of urea from 300 to 450 kg/ha enhanced the photosynthesis of leaves, promoted the synthesis of soluble sugars, thus providing sufficient nutrition for the growth and development of grape fruits.

The purpose of this study was to identify and analyze HXK gene family in grape. Based on the HXK(hexokinase) gene registration sequence of Arabidopsis thaliana and maize, four HXK genes were identified from the whole genome of Vitis vinifera, namd as VvHXK1, VvHXK2, VvHXK3 and VvHXK4. The bioinformatics analysis were performed on these four genes. The results of protein physicochemical properties showed that the number of amino acids encoded by the four genes was between 412 aa and 524 aa, of which VvHXK4 was a basic amino acid, and the other three were acidic amino acids. In addition, subcellular localization analysis indicated that all four VvHXK genes were localized in the cytoplasm. VvHXK4 was mainly localized in chloroplasts, and VvHXK2 might appear in mitochondria and nucleus. Meanwhile, the secondary structure prediction of the proteins found that the proteins encoded by the four VvHXK genes were mainly α-helix. The structural analysis of exons indicated that VvHXK1 contained 10 exons, and others contained 9 exons, showing that the VvHXK genes were relatively conserved; The initiation of cis-acting element analysis indicated that four VvHXK genes contained ABA response elements, MYB binding region and WRKY transcription factors, but no low temperature response component were found in VvHXK4. Phylogenetic tree analysis showed that the homology relationship between VvHXK1 and Arabidopsis AtHXK1 was the closest, indicating that it not only had the functions of regulating growth and development and affecting the growth of roots, but also had the effect of accelerating aging. The homology relationship between VvHXK2 and potato StHXK1 was recent, showing that it could increase the glucose phosphorylation activity in leaf extract or chloroplast. The homology relationship between VvHXK3 and potato StHXKRP1 was the closest, suggesting that VvHXK2 might be able to induce the expression of sugar in leaves. The homology relationship between VvHXK4 and spinach SoHXK1 was recent, which was important for studying the glucose metabolism of grapes. Fluorescence quantitative analysis showed that VvHXK2, VvHXK3 and VvHXK4 had the highest relative expression under the treatment of 20 g/L glucose, which was the most significant for glucose phosphorylation, and low expression under the treatment of fructose and sucrose. This is important for studying the sugar metabolism of grapes.

In order to define the characteristic aromatic components of each grape flavor type，we detected and analyzed the aromatic components of the berries from fifteen grape varieties (include 2 non-aromatic, 5 strawberry aromatic,8 muscat aromatic) using solid phase micro-extraction (SPME) coupled with gas chromatography-mass spectrometry (GC-MS).Results showed that the primary components of non-aromatic grape Moldova, Victoria is aldehydes. The relative content of 2-hexenal was the highest among aroma substances;The aroma substances of strawberry aromatic grape Shenfeng, Kyoho, Hupei No.2, Liaofeng, Shuofeng were mainly aldehydes, alcohols and esters; The aromatic substances of muscat aromatic grape were mainly aldehydes and terpenes. The main aldehydes are 2-hexenal and 1-hexanal.Terpenes were mainly linalool, dihydrolinalool, alpha-terpineol, nerol and geraniol. There were differences in aroma substances and substance contents among different grape varieties. This study can provide a theoretical basis for the research of grape aroma mechanism, breeding and promotion in the later stage.