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.

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.

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.

The role of the auxin-response genes MdGH3.1 and MdSAUR36 in bitter pit formation in apple

Apples often exhibit bitter pits in response to metabolic disorders during ripening and storage;however, the mechanisms underlying the bitter pit(BP) development remain unclear. Here, metabolome and transcriptome analyses were performed to investigate BP pulp of 'Fuji'. Two auxin-response genes, MdGH3.1 and MdSAUR36, were screened. Their expression as well as the auxin content in BP pulp were found to be higher than those in healthy pulp(P < 0.01). In the field, excess CO(NH2)2increased the incidence of BP. Moreover, the auxin content and MdGH3.1 expression increased in apples after nitrogen fertilization. On Day 30 before harvest, the two genes were transiently transferred to the fruit, and 20.69% and 23.21% of BP fruits were harvested. After 10 μmol·L-1auxin was infiltrated at low pressure into postharvest fruit, the increase in MdGH3.1 expression occurred earlier than that in MdSAUR36. MdGH3.1 increased the expression of MdSAUR36, but MdSAUR36 did not increase expression of MdGH3.1. Therefore, we suggest that MdGH3.1 acts upstream of MdSAUR36 during BP formation and that these genes induce BP formation by regulating auxin and phenylpropanoid biosynthesis.

Silencing of early auxin responsive genes MdGH3-2/12 reduces the resistance to Fusarium solani in apple

Apple replant disease(ARD)has led to severe yield and quality reduction in the apple industry.Fusarium solani(F.solani)has been identified as one of the main microbial pathogens responsible for ARD.Auxin(indole-3-acetic acid,IAA),an endogenous hormone in plants,is involved in almost all plant growth and development processes and plays a role in plant immunity against pathogens.Gretchen Hagen3(GH3)is one of the early/primary auxin response genes.The aim of this study was to evaluate the function of MdGH3-2 and MdGH3-12 in the defense response of F.solani by treating MdGH3-2/12 RNAi plants with F.solani.The results show that under F.solani infection,RNAi of MdGH3-2/12 inhibited plant biomass accumulation and exacerbated root damage.After inoculation with F.solani,MdGH3-2/12 RNAi inhibited the biosynthesis of acid-amido synthetase.This led to the inhibition of free IAA combining with amino acids,resulting in excessive free IAA accumulation.This excessive free IAA altered plant tissue structure,accelerated fungal hyphal invasion,reduced the activity of antioxidant enzymes(SOD,POD and CAT),increased the reactive oxygen species(ROS)level,and reduced total chlorophyll content and photosynthetic ability,while regulating the expression of PR-related genes including PR1,PR4,PR5 and PR8.It also changed the contents of plant hormones and amino acids,and ultimately reduced the resistance to F.solani.In conclusion,these results demonstrate that MdGH3-2 and MdGH3-12 play an important role in apple tolerance to F.solani and ARD.

Transcriptional regulation of MdPIN7 by MdARF19 during gravityinduced formation of adventitious root GSA in self-rooted apple stock

Self-rooted apple stock is widely used for apple production.However,the shallowness of the adventitious roots in self-rooted apple stock leads to poor performance in the barren orchards of China.This is because of the considerable difference in the development of a gravitropic set-point angle(GSA)between self-rooted apple stock and seedling rootstock.Therefore,it is crucial to study the molecular mechanism of adventitious root GSA in self-rooted apple stock for breeding self-rooted and deep-rooted apple rootstock cultivars.An apple auxin response factor MdARF19 functioned to establish the adventitious root GSA of self-rooted apple stock in response to gravity and auxin signals.MdARF19 bound directly to the MdPIN7 promoter,activating its transcriptional expression and thus regulating the formation of the adventitious root GSA in 12-2 self-rooted apple stock.However,MdARF19 influenced the expression of auxin efflux carriers(MdPIN3 and MdPIN10)and the establishment of adventitious root GSA of self-rooted apple stock in response to gravity signals by direct activation of MdFLP.Our findings provide new information on the transcriptional regulation of MdPIN7 by auxin response factor MdARF19 in the regulation of the adventitious root GSA of self-rooted apple stock in response to gravity and auxin signals.

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.

Effect of Different Raw Material Processing Methods on the Quality of Apple Distilled Spirits

In this study,the effect of yeast strains(X16 and RMS2),fruit seed,and pectinase on the quality of apple distilled spirits were investigated with apple as material.The results showed that pectinase shortened the fermentation period.The strain X16,fruit seed remaining,and pectinase addition groups had higher yields of crude distilled spirits than the strain RMS2,fruit seed removal,and without pectinase groups,respectively.Regarding the first-grade distilled spirits quality,the X16 group had higher content of total acids,total esters,higher alcohols,and alcohol content than the RMS2 group;the group with fruit seeds had higher total acids but lower alcohol content and total esters than the group with fruit seed removal;the group with pectinase addition had higher total acids and alcohol content than the group without pectinase addition.Regarding the second-grade distilled spirits quality,the X16 group had higher total acids,total esters,and alcohol content than the RMS2 group;the group without fruit seeds had higher alcohol content than the group with fruit seeds;the group with pectinase addition showcased higher total acids but lower alcohol content than the group without pectinase addition.In summary,yeast strains and pectinase affected the quality of apple distilled spirits,and strain X16 was more suitable for brewing apple distilled spirits.Pectinase affected the quality of apple distilled spirits by affecting fermentation rate and temperature.

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.

Isolation of Trichoderma virens 6PS-2 and its effects on Fusarium proliferatum f. sp. Malus domestica MR5 related to apple replant disease in China

Apple replant disease(ARD) is primarily caused by biotic factors that seriously inhibits the development of apple industry. Therefore, the use of biological control measures to inhibit the main pathogens(such as Fusarium spp.) that cause ARD is of great significance to the sustainable development of the apple industry. Trichoderma virens 6PS-2, which exhibited antagonism toward a variety of pathogens, was screened from the rhizosphere soils of healthy apple trees(Malus robusta) in different replanted orchards in the Yantai and Zibo Cities, Shandong Province, China. Its fermentation extract inhibited the growth of pathogenic Fusarium proliferatum f. sp. Malus domestica MR5, which was proportional to the concentration. These substances also increased the hairy root volume and growth of Arabidopsis thaliana lateral roots. The phenotype of Malus hupehensis seedlings and microbial community structure in rhizosphere soils in greenhouse experiment using Highthroughput sequencing were analyzed, and the field experiment with grafted apple trees were used for further verification. Compared with the application of potato dextrose broth(PDB) medium, application of 6PS-2 spore suspension directly to replanted soils could improve the growth of M. hupehensis seedlings as well as the elongation of grafted apple trees. Concomitant decreases in the gene copy number of Fusarium and increases in the culturable bacteria/fungi were also observed in the greenhouse and field experiments. The abundance of Trichoderma,Bacillus, and Streptomyces increased significantly, but that of Fusarium, Pseudarthrobacter, and Humicola decreased. The content of esters, phenols,furans, and amino acids in root exudates of M. hupehensis seedlings increased, which significantly inhibited the multiplication of Fusarium, but was positively correlated with Bacillus and Trichoderma. In summary, T. virens 6PS-2 not only directly inhibits the activity of pathogenic Fusarium but also secrets secondary metabolites with antifungal and growth-promoting potential. In addition, 6PS-2 spore suspension can also promote the growth of plants to a certain extent, and change the soil microbial community structure of rhizosphere soils. It is believed that T. virens 6PS-2 has the potential for the alleviation of apple replant disease(ARD) in China.

ALL ABOUT APPLES!

Crunch!Apples are a sweet and delicious fruit.There are over 7,500 different kinds!The fruit is usually red,but it can be green or other colors.Applesaren't expensive.They make a great snack.You can eat a whole apple.You can slice it,too.They're also good with other foods.Do you like apples?

Effect of Metaxenia on Volatile Compounds in Bagged Apple Fruit of Fuji

In the present study, the effect of pollen grain sources on volatile com- pounds in bagged apple fruit of Yan Fuji 3, grown at two experiment sites and pol- linated by pollens of other five cultivars was evaluated, to select appropriate poll- enizer cultivars for Fuji, and to provide a theoretical basis for improving the flavor and aroma of bagged apple fruit of Fuji. The results proved the significant metaxe- nia effect of the five pollenizer cultivars on volatile compounds of bagged apple fruit of Fuji. The numbers of both volatile compounds and characteristic aroma com- pounds in bagged apple fruit pollinated by Golden Delicious were larger and those of the fruit pollinated by Gala were smaller at both experiment sites. However, there were differences in the numbers of volatile compounds and characteristic aroma compounds among the fruits pollinated by Starkrimson, Jinxiuhaitang and Ruby at both experimental sites. The differences were caused by 12 compounds that only existed at a low level in certain samples. All of them were not the aroma contribu- tors except ethyl 2-methylbutyrate. The content of alcohols in bagged Fuji fruit was largely increased by Starkrimson and Ruby pollination; the content of aldehydes was obviously increased by Golden Delicious, Starkrimson and Gala pollination; the con- tents of esters and terpenes were significantly increased by Golden Delicious, Starkrimson and Ruby pollination. Jinxiuhaitang pollination showed no significant ef- fects for improving the content of four categories of volatile compounds. The metax- enia of the pollenizer cultivars on volatile compounds in bagged Fuji fruit was stable at the two experimental sites.

Isolation and Identification of Anthracnose Pathogen from Fruit of Red Fuji Apple

The fruits of Red Fuji apple with anthracnose symptoms were collected and submitted to tissue isolation and culture. One strain of anthracnose pathogen （numbered as Acgl） was obtained, and it was identified by both morphological and molecular biological methods. According to the morphological characteristics of the colony and conidia and the results of rDNA-ITS sequence analysis, the Acgl strain was identified as Colletotrichum gloeosporioides.

Analysis on Fuji Apple Tree Structures and Related Factors under Different Pruning Modes

To determine the correlations between the tree structuresof Fuji apple with different pruning modes and each factor, the data about 3 tree structures which were free spindle short shoot, free spindle long shoot and slenderspindle short shoot in Xingtang County of Hebai Province were investigated, then by SPSS anal- ysis, the correlations between the taperingness and each growth factor of inserted small branch were compared. The results showed that the taperingness of central trunk of free spindle dwarf-shoot Fuji apple treeshad negative correlations with each factor of inserted small branch, while the taperingness of central trunk of free spin- dle long-shoot Fuji apple treeshad positive correlations with each factor of inserted small branch, the taperingness of central trunk of slenderspindle short-shootFuji ap- ple treeshad negative correlation with total thickness of inserted small branch, but had positive correlations with other factors. This study can provide a scientifictheo- retical basis for the pruning technology of high-density planting trees grafting by dwarfing self-rooted rootstock.

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 Two Different Apple Dwarf Stocks on Juvenile Phase of Fuji Seedlings

The effects of apple dwarf stocks P22 and SH3 on the juvenile phase of Fuji seedlings were studied. The results showed that the juvenile phase could be shortened by more than two years when 2-year seedlings were grafted on the dwarf stocks. The effects of P22 and SH3 were significantly different. Grafted on P22, the flowering node was less, the first flowing site was lower, and the juvenile phase was shorter than SH3. With the two stocks, the average first flowering site was more than 200 cm, the average flowering node was more than 100, and the lateral branch top bud was more than 50% in all first flowing seedlings.

Correlation Analysis between Quality Characteristics and Fruit Mineral Element Contents in 'Fuji' Apples

[Objective] The aims were to explore the relationship between the contents of fruit mineral elements and quality features of the ＇Fuji＇ apple, screen major mineral elements of the fruit affecting fruit quality features, and set up optimum proposals of fruit mineral elements for good fruit qualities, so as to provide a theoretical basis for the reasonable orchard soil and foliar fertilizer applications to increase fruit quality and reduce the physiological diseases related to the ＇Fuji＇ apple. [Methods] The fruit mineral elements and quality indicators of ＇Fuji＂ apples were in- vestigated and analyzed, which were collected from the 153 commercial apple or- chards of ＂Fuji＇ apple located in 51 counties. The variable importance for projection （VlP） of partial least squares regression （PLS） method was used to analyze the model effect and weight analysis impact of the fruit mineral element contents to fruit quality, screen out major factors of fruit mineral elements influencing the different fruit qualities, and set up the regression equation of the fruit qualities and major fruit mineral elements. Linear programming was used to obtain optimum proposals of the fruit mineral elements to achieve good ＇Fuji＇ apple qualities. [Results] The mineral elements content and quality characteristics in ＂Fuji＇ apple fruit had great differences in the different produce regions in which the maximum content of nitro- gen, iron, zinc and boron in the ＇Fuji＇ fruit were12.06, 6.17, 7.7, and 10.08 times greater than the minimum respectively, and the differences for titratable acid and the SSC/TA ratio were 2.33 and 2.16 times respectively. The correlation analysis between the fruit mineral element contents and qualities showed that the nitrogen content of fruit had a significantly negative correlation with the soluble solid content, SSC/TA ratio and red color area, while the calcium and iron contents in the fruit were in significantly positive correlation with the soluble solid content and SSC/TA ratio. The model effect and weight analysis showed that the content of nitrogen and iron in the fruit had a greater influence on the integral fruit quality, followed by phosphorus, potassium and calcium. The variable importance for projection （VlP） technology of PLS found that the mean fruit weight was primarily affected by nitro- gen, phosphorus and potassium, and the soluble solid was primarily affected by ni- trogen, calcium and iron, while the red color area was primarily affected by nitro- gen, potassium, calcium, iron and zinc. The regression equation between fruit quality and mineral element contents showed that the mean fruit weight had a greater pos- itive effect coefficient with the content of phosphorus and potassium, and a greater negative effect coefficient with the content of nitrogen in the fruit. Moreover, the sol- uble solid had the largest negative effect coefficient with nitrogen and largest posi- tive effect coefficient with calcium and iron in the fruit. [Conclusion] The maximum content of soluble solid and titratable acid were 1.5 times more than the minimum, and nitrogen, iron, zinc and boron were 6 times more than in the ＇Fuji＇ apple fruit in the different produce regions. Therefore, it is a key technological measure to improve the overall qualities of the ＂Fuji＇ apple by decreasing the content of nitrogen, and increasing the contents of iron, phosphorus, potassium and calcium in the fruit.

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.