Comparative transcriptome analysis reveals key genes and pathways in response to Alternaria alternata apple pathotype infection

Apple leaf spot,caused by the Alternaria alternata apple pathotype(AAAP),is an important fungal disease of apple.To understand the molecular basis of resistance and pathogenesis in apple leaf spot,the transcriptomes of two apple cultivars‘Hanfu'(HF)(resistant)and‘Golden Delicious'(GD)(susceptible)were analyzed at 0,6,18,24 and 48 h after AAAP inoculation by RNA-Seq.At each time point,a large number of significantly differentially expressed genes(DEGs)were screened between AAAP-inoculated and uninoculated apple leaves.Analysis of the common DEGs at four time points revealed significant differences in the resistance of‘HF'and‘GD'apple to AAAP infection.RLP,RNL,and JA signal-related genes were upregulated in both cultivars to restrict AAAP development.However,genes encoding CNLs,TNLs,WRKYs,and AP2s were only activated in‘HF'as part of the resistance response,of which,some play major roles in the regulation of ET and SA signal transduction.Further analysis showed that many DEGs with opposite expression trends in the two hosts may play important regulatory roles in response to AAAP infection.Transient expression of one such gene MdERF110 in‘GD'apple leaves improved AAAP resistance.Collectively,this study highlights the reasons for differential resistance to AAAP infection between‘HF'and‘GD'apples which can theoretically assist the molecular breeding of disease-resistant apple crops.

Overexpression of the transcription factor MdWRKY115 improves drought and osmotic stress tolerance by directly binding to the MdRD22 promoter in apple

Abiotic stress reduces plant yield and quality.WRKY transcription factors play key roles in abiotic stress responses in plants,but the molecular mechanisms by which WRKY transcription factors mediate responses to drought and osmotic stresses in apple(Malus×domestica Borkh.)remain unclear.Here,we functionally characterized the apple GroupⅢWRKY gene MdWRKY115.qRT-PCR analysis showed that MdWRKY115 expression was up-regulated by drought and osmotic stresses.GUS activity analysis revealed that the promoter activity of MdWRKY115 was enhanced under osmotic stress.Subcellular localization and transactivation assays indicated that MdWRKY115 was localized to the nucleus and had a transcriptional activity domain at the N-terminal region.Transgenic analysis revealed that the overexpression of MdWRKY115 in Arabidopsis plants and in apple callus markedly enhanced their tolerance to drought and osmotic stresses.DNA affinity purification sequencing showed that MdWRKY115 binds to the promoter of the stress-related gene MdRD22.This binding was further verified by an electrophoretic mobility shift assay.Collectively,these findings suggest that MdWRKY115 is an important regulator of osmotic and drought stress tolerance in apple.

Biochar alleviates apple replant disease by reducing the growth of Fusarium oxysporum and regulating microbial communities

Apple replant disease(ARD)negatively affects plant growth and reduces yields in replanted orchards.In this study,biochar was applied to apple replant soil with Fusarium oxysporum.Our aim was to investigate whether biochar could promote plant growth and alleviate apple replant disease by reducing the growth of harmful soil microorganisms,changing soil microbial community structure and improving the soil environment.This experiment included five treatments:apple replant soil(CK),methyl bromide fumigation apple replant soil(FM),replant soil with biochar addition(2%),replant soil with F.oxysporum spore solution(8×10^(7)spores·mL^(-1)),and replant soil with biochar and F.oxysporum spore solution addition.Seedling biomass,the activity of antioxidant enzymes in the leaves and roots,and soil environmental variables were measured.Microbial community composition and community structure were analyzed using 16SrDNA and ITS2 gene sequencing.Biochar significantly reduced the abundance of F.oxysporum and increased soil microbial diversity and richness.Biochar also increased the soil enzyme activities(urease,invertase,neutral phosphatase,and catalase),the biomass(plant height,fresh weight,dry weight)and the activity of antioxidant enzymes(superoxide dismutase,peroxidase,and catalase).The root indexes of apple seedlings was also increased in replant soil by biochar.In sum,biochar promoted the growth of plants,improved the replant soil environment,and alleviated apple replant disease.

Improved multi-scale inverse bottleneck residual network based on triplet parallel attention for apple leaf disease identification

Accurate diagnosis of apple leaf diseases is crucial for improving the quality of apple production and promoting the development of the apple industry. However, apple leaf diseases do not differ significantly from image texture and structural information. The difficulties in disease feature extraction in complex backgrounds slow the related research progress. To address the problems, this paper proposes an improved multi-scale inverse bottleneck residual network model based on a triplet parallel attention mechanism, which is built upon ResNet-50, while improving and combining the inception module and ResNext inverse bottleneck blocks, to recognize seven types of apple leaf(including six diseases of alternaria leaf spot, brown spot, grey spot, mosaic, rust, scab, and one healthy). First, the 3×3 convolutions in some of the residual modules are replaced by multi-scale residual convolutions, the convolution kernels of different sizes contained in each branch of the multi-scale convolution are applied to extract feature maps of different sizes, and the outputs of these branches are multi-scale fused by summing to enrich the output features of the images. Second, the global layer-wise dynamic coordinated inverse bottleneck structure is used to reduce the network feature loss. The inverse bottleneck structure makes the image information less lossy when transforming from different dimensional feature spaces. The fusion of multi-scale and layer-wise dynamic coordinated inverse bottlenecks makes the model effectively balances computational efficiency and feature representation capability, and more robust with a combination of horizontal and vertical features in the fine identification of apple leaf diseases. Finally, after each improved module, a triplet parallel attention module is integrated with cross-dimensional interactions among channels through rotations and residual transformations, which improves the parallel search efficiency of important features and the recognition rate of the network with relatively small computational costs while the dimensional dependencies are improved. To verify the validity of the model in this paper, we uniformly enhance apple leaf disease images screened from the public data sets of Plant Village, Baidu Flying Paddle, and the Internet. The final processed image count is 14,000. The ablation study, pre-processing comparison, and method comparison are conducted on the processed datasets. The experimental results demonstrate that the proposed method reaches 98.73% accuracy on the adopted datasets, which is 1.82% higher than the classical ResNet-50 model, and 0.29% better than the apple leaf disease datasets before preprocessing. It also achieves competitive results in apple leaf disease identification compared to some state-ofthe-art methods.

Overexpression of auxin/indole-3-acetic acid gene MdIAA24 enhances Glomerella leaf spot resistance in apple(Malus domestica)

Auxin is throughout the entire life process of plants and is involved in the crosstalk with other hormones,yet its role in apple disease resistance remains unclear.In this study,we investigated the function of auxin/indole-3-acetic acid(IAA)gene Md IAA24 overexpression in enhancing apple resistance to Glomerella leaf spot(GLS)caused by Colletotrichum fructicola(Cf).Analysis revealed that,upon Cf infection,35S::Md IAA24 plants exhibited enhanced superoxide dismutase(SOD)and peroxidase(POD)activity,as well as a greater amount of glutathione(reduced form)and ascorbic acid accumulation,resulting in less H_(2)O_(2)and superoxide anion(O_(2)^(-))in apple leaves.Furthermore,35S::Md IAA24 plants produced more protocatechuic acid,proanthocyanidins B1,proanthocyanidins B2 and chlorogenic acid when infected with Cf.Following Cf infection,35S::Md IAA24 plants presented lower levels of IAA and jasmonic acid(JA),but higher levels of salicylic acid(SA),along with the expression of related genes.The overexpression of Md IAA24 was observed to enhance the activity of chitinase andβ-1,3-glucanase in Cfinfected leaves.The results indicated the ability of Md IAA24 to regulate the crosstalk between IAA,JA and SA,and to improve reactive oxygen species(ROS)scavenging and defense-related enzymes activity.This jointly contributed to GLS resistance in apple.

Potassium alleviated high nitrogen-induced apple growth inhibition by regulating photosynthetic nitrogen allocation and enhancing nitrogen utilization capacity

There is a close relationship between potassium(K)and nitrogen(N).However,the roles of K under high N conditions remain unclear.Using a hydroponics approach,we monitored the morphological,physiological,and molecular changes in M9T337 apple(Malus domestica)rootstocks under different nitrate(10 and 30 mmol·L^(-1)NO_(3)^(-))and K supply(0.5,6,10,and 20 mmol·L_(-1)K^(+))conditions.Results revealed that high nitrate inhibited the root growth of M9T337 rootstocks,downregulated the expressions of K transporter genes(MdPT5,MdHKT1,and MdATK1),and reduced the net NO3-and K+influx at the surface of roots,thereby resulting in an N/K imbalance in rootstocks.Further investigation showed that 10 mmol·L^(-1)K increased the activity of N metabolic enzymes(NR,GS,NiR,and GOGAT),upregulated the expressions of genes related to nitrate uptake and transport(MdNRT1.1,MdNRT1.2,MdNRT1.5,and MdNRT2.4),promoted15N transport from the roots to the shoots,optimized leaf N distribution,and improved photosynthetic N utilization efficiency under high nitrate conditions.These results suggest that the negative effects of high nitrate may be related to the N/K imbalance and that reducing N/K in plants by increasing K supply level can effectively alleviate the inhibition of N assimilation by high nitrate stress.

HATCH评分、APPLE评分在心房颤动患者射频消融术后复发的预测价值研究

目的探讨HATCH评分、APPLE评分在心房颤动(房颤)患者射频消融术后复发的预测价值。方法选取2018年5月至2020年6月于秦皇岛市第一医院心血管内科行射频消融术治疗的105例房颤患者的临床资料进行回顾性分析,观察手术时间、X线曝光时间、消融成功率及复发率;且根据术后3个月后是否出现持续时间≥30 s的快速性房性心律失常分为复发组(n=36)和未复发组(n=69),分析影响复发的危险因素,评估HATCH评分、APPLE评分及两项联合评估房颤患者术后复发的AUC值、敏感度、特异度。结果105例房颤患者均顺利完成射频消融术,消融成功率为65.71%,复发率为34.29%。复发组与未复发组在性别、年龄、病程、合并高血压、糖尿病、心力衰竭、估算的肾小球滤过率(eGFR)、持续性房颤、左心室大小、左室射血分数(LVEF)、右室前后径、升主动脉内径、肺动脉主干内径、室内隔厚度、左室后壁厚度、主动脉瓣口血流速度、肌酐等资料比较中,差异均无统计学意义(P>0.05);而在左心房大小、E峰、HATCH评分、APPLE评分等比较中,差异具有统计学意义(P<0.05)。二元Logistic回归分析显示,左心房大小、E峰、HATCH评分、APPLE评分是影响手术治疗后复发的独立危险因素(P<0.001)。ROC曲线分析显示,HATCH评分、APPLE评分及两项联合评估房颤患者术后复发的AUC值分别为(0.747、0.807、0.927,P<0.05);敏感度分别为44.40%、69.40%、94.40;特异度分别为91.30%、85.50%、84.10%。结论HATCH评分、APPLE评分预测房颤患者射频消融术后复发具有一定价值,且两项联合有利于提高敏感度,能为识别术后复发提供指导。

高龄患者围术期APPLE-MDT系统设计与应用

目的以患者为中心,以“全人、个体化”诊疗为目标,采用多学科协作围术期全程管理模式,保障高龄手术患者的医疗安全。方法建设高龄患者围术期APPLE-MDT系统,通过多学科专家评估量表,自动生成评估报告,指导临床医生对患者术前、术中、术后、远期进行有针对性的干预治疗。结果APPLE-MDT管理模式应用于高龄骨科患者围术期,初步实现人工智能辅助决策,高龄患者例数逐年增加,术前住院日和住院天数减少,术后并发症发生率降低到2.4%,可手术率提高到90%,术后死亡率为0。结论高龄患者围术期APPLE-MDT系统辅助建立了高龄患者围术期全数据链数据库,构建了高龄患者围术期新路径管理,体现了先进的医疗管理理念,推动了高质量科研产出。

Effects of water stress on quality and sugar metabolism in'Gala'apple fruit

Sugar plays an important role in apple fruit development,appearance and quality as well as contributing to a plant’s water stress response.Trehalose and the trehalose biosynthetic metabolic pathways are part of the sugar signaling system in plants,which are important regulator of water stress response in apple.The effect of water stress treatments applied to apple trees and the corresponding effects of ABA on developmental fruit quality were examined for indicators of fruit quality during fruit development.The results indicated that the severe water stress treatment(W2)occurring after the last stage of fruit cell division caused a decrease in the color and size of fruit.The moderate water stress(W1)occurring after the last stage of fruit cell enlargement(S2)caused an increase in the content of fructose and sorbitol while the apple fruit shape was not affected.These changes in sugar are related to the activity of sugar metabolic enzymes.While the enzymatic activity of vacuolar acid invertase(vAINV)was higher,that of sucrose-phosphate synthase(SPS)was lower in water stress treated fruit throughout the developmental period.This indicates that enhanced sucrose degradation and reduced sucrose synthesis leads to an overall reduced sucrose content during times of drought.Thus,water stress reduced sucrose content.Whereas the content of endogenous trehalose and ABA were the highest in water stress treated fruit.A moderate water stress(W1)imposed on apple trees via water restriction(60%–65%of field capacity)after the fruit cell enlargement phase of fruit development yielded sweeter fruit of higher economic value.

Extracellular and intracellular infection of Botryosphaeria dothidea and resistance mechanism in apple cells

Apple ring rot,which is caused by Botryosphaeria dothidea,severely affects apple production.The mechanisms employed in apple cells against B.dothidea remain unknown.In this research,the pathogen infection mode and the relationship between cell death and disease resistance in‘Fuji’/B.dothidea interaction pathosystem were investigated.By using transmission electron microscopy(TEM),our research showed that the pathogen infects apple cells both intracellularly and extracellularly.However,compared with that in immature fruit,the incidence of hyphae in the interior of mature apple fruit cells increased dramatically,suggesting that cell wall-mediated penetration resistance could be important in apple resistance against B.dothidea.TEM ultrastructural characterization identified the nuclear morphology of programmed cell death induction in both apple fruit and callus cells under B.dothidea infection.Overexpression of MdVDAC2(MDP0000271281),which encodes an outer-membrane localized anion channel protein in mitochondria,significantly promoted cell death under B.dothidea infection and simultaneously inhibited pathogen infection,suggesting that cell death represents a disease resistance mechanism in apple against B.dothidea infection.Furthermore,BdCatalase(KAF4307763),a cytochromeP450 family protein BdCYP52A4(KAF4300696),and subtilisin-domain containing proteinswere identified fromB.dothidea-secreted proteins,which suggested the potential involvement of active oxygen species and phytoalexins in combating B.dothidea infection and triggering or dampening apple resistance.Collectively,our research suggested that cell wall-mediated penetration resistance,programmed cell death machinery and microbial effector-interrelated signaling were among strategies recruited in apple to combat B.dothidea.The current research laid the foundation for further investigations into resistance mechanisms in apple.

Comparison of transcriptome and metabolome analysis revealed differences in cold resistant metabolic pathways in different apple cultivars under low temperature stress

Freezing injury in winter is an important abiotic stress that seriously affects plant growth and development.Deciduous fruit trees resist freezing injury by inducing dormancy.However,different cultivars of the same species have different cold resistance strategies.Little is known about the molecular mechanism of apple trees in response to freezing injury during winter dormancy.Therefore,in this study,1-year-old branches of the cold-resistant cultivar‘Hanfu’(HF)and the cold-sensitive cultivar‘Changfuji No.2’(CF)were used to explore their cold resistance through physiological,biochemical,transcriptomics,and metabolomics analyses.Combining physiological and biochemical data,we found that HF had a stronger osmotic regulation ability and antioxidant enzyme activity than CF,as well as stronger cold resistance.The functional enrichment analysis showed that both cultivars were significantly enriched in pathways related to signal transduction,hormone regulation,and sugar metabolism under freezing stress.In addition,the differentially expressed genes(DEGs)encoding galactinol synthase,raffinose synthase,and stachyose synthetase in raffinose family oligosaccharides(RFOs)metabolic pathways were upregulated in HF,and raffinose and stachyose were accumulated,while their contents in CF were lower.HF accumulated 4-aminobutyric acid,spermidine,and ascorbic acid to scavenge reactive oxygen species(ROS).While the contents of oxidized glutathione,vitamin C,glutathione,and spermidine in CF decreased under freezing stress,consequently,the ability to scavenge ROS was low.Furthermore,the transcription factors apetala 2/ethylene responsive factor(AP2/ERF)and WRKY were strongly induced under freezing stress.In summary,the difference in key metabolic components of HF and CF under freezing stress is the major factor affecting their difference in cold resistance.The obtained results deepen our understanding of the cold resistance mechanism in apple trees in response to freezing injury during dormancy.

YOLO-RD-Apple果园异源图像遮挡果实检测模型

为探索在自然苹果园环境中对高度遮挡果实进行机器人自动化采摘,提出基于RGB与Depth图像双输入的YOLO-RD-Apple果园异源图像遮挡果实检测模型。使用轻量化的MobileNetV2并在此基础上设计的更加轻量化的MobileNetV2-Lite分别作为RGB和Depth图像的特征提取器,保证特征提取能力的同时降低网络的计算量;将CSPNet与深度可分离卷积结合SE注意力模块,提出全新的SE-DWCSP3模块对PANet结构进行改进,提升网络对于残缺苹果目标的特征提取能力;引入SoftNMS算法替代一般NMS算法,以减少对密集目标的错误抑制现象,降低被遮挡苹果的漏检率。实验结果表明,YOLO-RD-Apple在自然遮挡苹果数据集上性能优秀,在测试集上的AP值达到93.1%,较YOLOv4提升了1.4%,参数量则降低了70%,在GPU(V100)的检测速度达到40.5FPS,较YOLOv4速度提升了12.5%,在检测精度和速度上均有不同程度地提升,同时降低了网络参数量,更加适用于实际果园苹果采摘场景。

Multivariate analysis between meteorological factor and fruit quality of Fuji apple at different locations in China

China has the largest apple planting area and total yield in the world, and the Fuji apple is the major cultivar, accounting for more than 70％ of apple planting acreage in China. Apple qualities are affected by meteorological conditions, soil types, nutrient content of soil, and management practices. Meteorological factors, such as light, temperature and moisture are key environmental conditions affecting apple quality that are difficult to regulate and control. This study was performed to determine the effect of meteorological factors on the qualities of Fuji apple and to provide evidence for a reasonable regional layout and planting of Fuji apple in China. Fruit samples of Fuji apple and meteorological data were investigated from 153 commercial Fuji apple orchards located in 51 counties of 11 regions in China from 2010 to 2011. Partial least-squares regression and linear programming were used to analyze the effect model and impact weight of meteorological factors on fruit quality, to determine the major meteorological factors influencing fruit quality attributes, and to establish a regression equation to optimize meteorological factors for high-quality Fuji apples. Results showed relationships between fruit quality attributes and meteorological factors among the various apple producing counties in China. The mean, minimum, and maximum temperatures from April to October had the highest positive effects on fruit qualities in model effect loadings and weights, followed by the mean annual temperature and the sunshine percentage, the temperature difference between day and night, and the total precipitation for the same period. In contrast, annual total precipitation and relative humidity from April to October had negative effects on fruit quality. The meteorological factors exhibited distinct effects on the different fruit quality attributes. Soluble solid content was affected from the high to the low row preface by annual total precipitation, the minimum temperature from April to October, the mean temperature from April to October, the temperature difference between day and night, and the mean annual temperature. The regression equation showed that the optimum meteorological factors on fruit quality were the mean annual temperature of 5.5-18°C and the annual total precipitation of 602-1121 mm for the whole year, and the mean temperature of 13.3-19.6°C, the minimum temperature of 7.8-18.5°C, the maximum temperature of 19.5°C, the temperature difference of 13.7°C between day and night, the total precipitation of 227 mm, the relative humidity of 57.5-84.0％, and the sunshine percentage of 36.5-70.0％ during the growing period （from April to October）.

Farmers’precision pesticide technology adoption and its influencing factors:Evidence from apple production areas in China

The research aimed to understand farmers’willingness to adopt(WTA)and willingness to pay(WTP)for precision pesticide technologies and analyzed the determinants of farmers’decision-making.We used a two-stage approach to consider farmers’WTA and WTP for precision pesticide technologies.A survey of 545 apple farmers was administered in Bohai Bay and the Loess Plateau in China.The data were analyzed using the double-hurdle model.The results indicated that 78.72%of respondents were willing to apply precision pesticide technologies provided by service organizations such as cooperatives and dedicated enterprises,and 69.72%were willing to buy the equipment for using precision pesticide technologies.The results of the determinant analysis indicated that farmers’perceived perceptions,farm scale,cooperative membership,access to digital information,and availability of financial services had significant and positive impacts on farmers’WTA precision pesticide technologies.Cooperative membership,technical training,and adherence to environmental regulations increased farmers’WTP for precision pesticide technologies.Moreover,nonlinear relationships between age,agricultural experience,and farmers’WTA and WTP for precision pesticide technology services were found.

Genome-wide identification and function analysis of the sucrose phosphate synthase MdSPS gene family in apple

Sucrose phosphate synthase(SPS)is a rate-limiting enzyme that works in conjunction with sucrose-6-phosphate phosphatase(SPP)for sucrose synthesis,and it plays an essential role in energy provisioning during growth and development in plants as well as improving fruit quality.However,studies on the systematic analysis and evolutionary pattern of the SPS gene family in apple are still lacking.In the present study,a total of seven MdSPS and four MdSPP genes were identified from the Malus domestica genome GDDH13 v1.1.The gene structures and their promoter cis-elements,protein conserved motifs,subcellular localizations,physiological functions and biochemical properties were analyzed.A chromosomal location and gene-duplication analysis demonstrated that whole-genome duplication(WGD)and segmental duplication played vital roles in MdSPS gene family expansion.The Ka/Ks ratio of pairwise MdSPS genes indicated that the members of this family have undergone strong purifying selection during domestication.Furthermore,three SPS gene subfamilies were classified based on phylogenetic relationships,and old gene duplications and significantly divergent evolutionary rates were observed among the SPS gene subfamilies.In addition,a major gene related to sucrose accumulation(MdSPSA2.3)was identified according to the highly consistent trends in the changes of its expression in four apple varieties(‘Golden Delicious’,‘Fuji’,‘Qinguan’and‘Honeycrisp’)and the correlation between gene expression and soluble sugar content during fruit development.Furthermore,the virus-induced silencing of MdSPSA2.3 confirmed its function in sucrose accumulation in apple fruit.The present study lays a theoretical foundation for better clarifying the biological functions of the MdSPS genes during apple fruit development.

Identification of the HAK gene family reveals their critical response to potassium regulation during adventitious root formation in apple rootstock

Adventitious root formation is a bottleneck during vegetative proliferation.Potassium(K^(+))is an essential macronutrient for plants.K^(+)accumulation from the soil and its distribution to the different plant organs is mediated by K^(+)transporters named K^(+)transporter(KT),K^(+)uptake(KUP),or high-affinity K^(+)(HAK).This study aimed to identify members of the HAK gene family in apples and to characterize the effects of K^(+)supply on adventitious root formation and on the expression of HAK genes and the genes that putatively control auxin transport,signaling,and cell fate during adventitious root formation.In this study,34 HAK genes(MdHAKs)were identified in the apple(Malus×domestica‘Golden Delicious’)genome.A phylogenetic analysis divided MdHAKs into four clusters(Ⅰ,Ⅱ,Ⅲ,andⅣ),comprising 16,1,4,and 13 genes,respectively.The syntenic relationships revealed that 62.5%of the total MdHAK genes arise from genomic duplication events.Chromosome location,domain structure,motif analysis,and physico-chemical characteristics were subsequently investigated.Furthermore,the application of K^(+)indicated the emergence of adventitious roots at 8 d and produced more adventitious roots at 16 d than the K^(+)-free control(CK)treatment.In addition,various MdHAKs showed root-specific expression in B9 apple rootstock stem cuttings and enhanced expression during the initiation and emergence stages of adventitious root formation in response to K^(+)treatment.Additionally,K^(+)treatment enhanced the expression levels of MdPIN1,MdPIN2,and MdAUX1.Further data indicated that a higher expression of MdWOX11,MdLBD16,and MdLBD29 and of cell cycle-related genes contributed to the auxin-stimulated adventitious root formation in response to K^(+).

MdDGK3-like as a negative regulator participates in ALA-induced PP2AC to promote stomatal opening in apple leaves

5-Aminolevulinic acid(ALA)can inhibit abscisic acid(ABA)-induced stomatal closure.However,the molecular mechanism is unclear.In this study,we found that ALA upregulated the MdPP2AC expression and PP2A activity in the apple(Malus domestica Borkh.cv.‘Fuji’)leaves.With the promoter of MdPP2AC as bait,a diacylglycerol kinase MdDGK3-like was selected by the Yeast One Hybrid(Y1H)from the cDNA library of the epidermis of apple leaves treated by exogenous ALA.Additional to binding the promoter of MdPP2AC,MdDGK3-like was found to inhibit the transcription activity of MdPP2AC promoter,while ALA significantly eliminated the role of MdDGK3-like.In tobacco leaves,MdDGK3-like was localized in the nucleus of stomatal guard cells.Therefore,MdDGK3-like might act as a transcription factor negatively regulating MdPP2AC expression and causing stomatal closure.To further identify MdDGK3-like functions,several transiently transgenic apple leaves(including overexpression and interference)were established.The results revealed that overexpression of MdDGK3-like promoted stomatal closure by increasing Ca^(2+)and H_(2)O_(2)and decreasing flavonol levels in the guard cells.Conversely,MdDGK3-like(i)led the stomatal opening with lower levels of Ca^(2+)and H_(2)O_(2)but higher flavonols.Based on these,we proposed a new hypothesis that ALA up-regulated MdPP2AC expression via negatively regulating the expression of MdDGK3-like to up-regulate PP2A expression and the enzyme activity,which improved the stomatal aperture.Since it was the first time that MdDGK3-like was showed to act as a transcription factor,the proposed model provided a new insight onto the mechanisms of ALA-induced stomatal opening.

Prediction of Apple Fruit Quality by Soil Nutrient Content and Artificial Neural Network

The effect of soil nutrient content on fruit yield and fruit quality is very important.To explore the effect of soil nutrients on apple quality we investigated 200 fruit samples from 40 orchards in Feng County,Jiangsu Province.Soil mineral elements and fruit quality were measured.The effect of soil nutrient content on fruit quality was analyzed by artificial neural network(ANN)model.The results showed that the prediction accuracy was highest(R2=0.851,0.847,0.885,0.678 and 0.746)in mass per fruit(MPF),hardness(HB),soluble solids concentrations(SSC),titratable acid concentration(TA)and solid-acid ratio(SSC/TA),respectively.The sensitivity analysis of the prediction model showed that soil available P,K,Ca and Mg contents had the greatest impact on the quality of apple fruit.Response surface method(RSM)was performed to determine the optimum range of the available P,K,Ca,and Mg contents in orchards In Feng County,which were 10∼20 mg⋅kg^(−1),170∼200 mg⋅kg^(−1),1000∼1500 mg⋅kg^(−1),and 80∼200 mg⋅kg^(−1),respectively.The research also concluded that improving the content of available P and available Ca in orchard soil was crucial to improve apple fruit quality in Feng County,Jiangsu Province.

Effect of Metaxenia on Sugars and Organic Acids and Taste Quality of Bagged Fuji Apple Fruits

In order to provide reference for the selection of appropriate pollination varieties facilitating the improvement of the flavor quality of bagged Fuji apple, the metaxenia effects of 5 pollination varieties on sugars, organic acids and flavor quality of bagged Fuji fruit were studied. The results showed that the 5 pollination varieties had significant metaxenia effects on sugars, organic acids and flavor quality of bagged Fuji fruit. For the sugar components, the fruits pollinated with Golden Delicious and Jinxiuhaitang showed higher glucose contents, and the fruit pollinated with Gala had the highest sucrose content; the fruits pollinated by Gala, Golden Delicious and Ruby exhibited higher malic acids, and the oxalic acid content and the succinic acid content were higher in the fruits pollinated with Jinxiuhaitang and Ruby ; and the fruit pollinated with Gala had the highest total sugar content, and the total acid content was higher in the fruits pollinated by Jinxiuhaitang and Ruby. The sugar-acid ratio was higher in the fruits pollinated with Gala and Golden Delicious, and the lowest in the fruit pollinated with Ruby. The senso- ry evaluation of fruits showed that the fruit pollinated with Ruby tasted sour-sweet, while the fruits pollinated with other 4 pollination varieties were sour and sweet moderately.

Analysis of Aroma Volatile Compounds in Fuji Apple Using SPME with Different Fiber Coatings

The aroma volatile compounds in Fuji apple were isolated by solid-phase microextraction( SPME),and detected by gas chromatography-mass spectrometry( GC-MS). The results showed that the optimal retention time of the heating up of capillary columns was 5 min. Of the seven fibers used in this work,DVB/CAR/PDMS was found to be the most suitable to extract the aroma volatile compounds in Fuji apple. A total number of 43 volatile compounds were identified according to their retention time on capillary columns and their mass spectra,including eleven esters,ten alcohols,ten aldehydes,seven alkenes,two anhydrides,one ketone,one phenol and one ether. This detection method will provide a new foundation for analysis of volatile compounds in apple were identified.