Genetic fingerprinting identifies apple (Malus domestica Borkh.) fruit cultivars in historic orchards on public lands

Genetic Resources and Crop Evolution - The National Historic Preservation Act of 1966 mandates that historic resources, including orchards, on federal lands be evaluated for potential significance....     

Investigations of aroma volatile biosynthesis under anoxic conditions and in different tissues of "Redchief Delicious" apple fruit (Malus domestica Borkh.)

Disks from different tissues were obtained from "Redchief Delicious" apple fruit (Malus domestica Borkh.) and analyzed for the ability to metabolize 1-pentanol as well as synthesize constitutive esters and alcohols under anoxic and aerobic conditions. The skin tissue displayed a greater capacity to synthesize pentanal, pentyl acetate, pentyl propionate, pentyl butyrate, and pentyl hexanoate than the hypanthial and carpellary tissues during incubation with 1-pentanol. With the exception of pentyl acetate and pentyl propionate biosynthesis, the hypanthial tissue synthesized these compounds at a higher rate than the carpellary tissue. Anoxia inhibited both constituent and 1-pentanol-derived ester biosynthesis. While anoxia inhibited ester biosynthesis, ethanol biosynthesis increased at a greater rate in tissue disks held under these conditions. Biosynthesis of 1-butanol, 2-methyl-1-butanol, and 1-hexanol was greater in tissue disks held in air during the first part of the measurement period and dropped off more rapidly than those transpiring in tissue disks held under anoxic conditions. The biosynthetic rates of all esters, both constituent and 1-pentanol-derived, increased as a result of air exposure. While hypoxic or anoxic conditions may promote ethanol synthesis, these conditions also appear to inhibit the formation of the ethanol-derived esters partially responsible for the off-flavor in apples attributed to ultralow O(2) controlled atmosphere storage.     

Re-Visiting Calcium Concentration and Distribution in Apple Fruit (Malus domestica Borkh.)

Distribution of calcium in the apple has been quantified using mineral analyses of fruit sections during the mid-1900s by various researchers. Most of the work was done either at harvest, or after storage. We used Proton-induced X-ray spectrometry (PIXE) and Scanning Electron Microscopy to correlate Ca distribution in apples to results obtained from mineral analyses (MA). For fruit at harvest transverse distribution of Ca in ‘Braeburn’ apples determined by PIXE confirmed similar analyses with MA. It was not possible to establish a significant relationship between the total Ca concentration determined by PIXE and soluble Ca concentration from MA in this paper, either at harvest, or 80 days after full bloom (dafb). A significant correlation was found between PIXE and SEM results 80 dafb.     

Postharvest variation in apple (Malus x domestica Borkh.) Flavonoids following harvest, storage, and 1-MCP treatment

The impact of 1-methylcyclopropene (1-MCP) on the synthesis and retention of flavonoid compounds during storage and ripening of red Delicious (Malus x domestica Borkh.) apples was investigated. Numerous anthocyanins, flavonols, flavan-3-ols, and a hydroxycinnamic acid from three different fruit harvest maturities were monitored after a 120 day storage and 1 week shelf life period using high-performance liquid chromatography/diode array detector analysis. The total flavonoid concentration was 5% greater in fruit treated with 1-MCP, whereas chlorogenic acid levels were 24% lower. All compounds analyzed increased in concentration during fruit harvest; however, the anthocyanins generally declined after storage, while chlorogenic acid levels increased. 1-MCP treatment resulted in the retention of anthocyanins in the latter stages of storage but did not affect the flavonols and flavan-3-ols. Chlorogenic acid biosynthesis from early and optimal fruit harvest maturities was greatly inhibited by 1-MCP during storage and the 1 week shelf life period. However, 1-MCP did not affect chlorogenic acid concentrations in late-harvested fruit. Results suggest that 1-MCP may inhibit the activity of phenylalanine ammonia-lyase and subsequent biosynthesis of flavonoid compounds. However, because very little postharvest biosynthesis of flavonoids occurs in apples, 1-MCP treatment may be useful for maintaining some of the intrinsic flavonoid levels of red Delicious apples, if applied at the proper harvest maturity.     

The role of root exudates in specific apple (Malus × domestica Borkh.) replant disease (SARD)

In an effort to improve our understanding of the specific apple replant disease (SARD), direct and indirect effects of phytohormones and related compounds (abscisic acid, 6‐benzyladenine, indole‐3‐acetic acid, 1‐naphthaleneacetic acid, and gibberellins GA₃ or GA₄) on root exudates of apple seedlings were evaluated as a potential mediating factor in the infection process. In the first type of experiments, radioactively labeled hormonal substances were applied to the stumps of decapitated apple‐seedlings and the occurrence of radioactivity in root exudates and their qualitative characterization were examined (direct influence). In another set of experiments, the effects of leaf‐applied plant growth regulators on the amount and composition of the predominant organic acids, carbohydrates, and amino acids/amides were studied. Cherry seedlings resistant to SARD and apple seedlings with dormant apical buds and, thus, not susceptible to infection were used for comparison. The results showed no differences in exudation of applied plant growth regulators between growing cherry and apple seedlings. Thus, a direct effect of plant hormones on the infection process is unlikely. However, leaf treatments with growth regulators, in particular with auxin‐type compounds and abscisic acid, increased exudation of alditols. This may indicate that plant hormones are, to some degree, indirectly involved in the infection process.     

Heavy metals mobility,sources, and risk assessment in soils and uptake by apple (Malus domestica Borkh.) leaves in urban apple orchards

The aim of the study was determine potential mobility, sources, and environmental impacts of heavy metals in soil of urban apple orchards using a sequential extraction procedure and multivariate analysis. The soil and apple (Malus domestica Borkh.) leaf samples were collected from apple orchards of urban areas in Çanakkale, Turkey. A sequential extraction procedure was employed to determine the binding forms of Cd, Cr, Cu, Ni, Pb, and Zn in soil samples. Sequential extraction procedure revealed that mobile fractions (acid soluble, reducible, and oxidizable) of Cd (60.2%) and Pb (56.8%) were higher than their immobile fractions. Such higher mobile fractions indicate the anthropogenic sources, and multivariate analysis results also supported the existence of such sources. The relationship between pH and mobile fractions of the metals (sum of acid soluble, reducible, and oxidizable) indicates that Cd and Pb become more mobile under decreasing pH conditions. Considering all metals, except for Cd, a strong relationship was observed between metal concentrations of apple leaves and mobile fractions, and plants were primarily able to uptake the metals in these fractions. Contamination factor and risk assessment code results together also revealed that Cd was highly mobile; retained less and had high risk for the environment.     

Lower crop load for Cv. Jonagold apples (Malus x domestica Borkh.) increases polyphenol content and fruit quality

The influence of crop load on fruit quality was investigated on 7-year-old slender spindle cv. Jonagold/M.9 apple trees. In mid June five different crop loads per tree were prepared by reducing the fruit number to average 30, 59, 104, 123, and 157 fruits per crown. The fruit from low-cropping trees had more red blush, a higher percentage of soluble solids in fruit flesh, and better flesh firmness in comparison to fruit from high-cropping trees. As the crop load decreased, the concentration of all phenolic compounds in the fruit samples (cortex plus skin) increased; concentrations of the most important individual fruit phenolics were also higher. When crop load fell from 157 to 30 fruits per crown, total polyphenols increased from an average of 1300 to 1680 mg/kg of fruit fresh weight (FW) (+29%), low molecular weight polyphenols increased from 1140 to 1570 mg/kg of FW (+38%), and high molecular weight polyphenols increased from 1740 to 2070 mg/kg of FW (+19%). The average increases in single polyphenols were even greater: chlorogenic acid (+82%); 4'-p-coumaroylquinic acid (+22%); catechin (+178%); and epicatechin (+71%). Ascorbic acid was not significantly dependent on crop load.     

Flowering phenology and the interrelations between phenological stages in apple trees (Malus domestica Borkh.) as influenced by the Nordic climate

In order to assess to what degree the ongoing global warming has affected flowering time of apple trees in the Nordic climate, we studied flowering phenology of trees in an orchard in Southern Norway. Correlation analysis of a 70-year (1946–2016) data series for flowering time of the cultivar ‘Gravenstein’ revealed a close relationship of the flowering phenophase with April–May heat accumulation (r?=??0.98). Over the last 50-year period, flowering was advanced by 16 days in response to the temperature rises. Similarly, on average for 12 cultivars of varying earliness, flowering was advanced by 9 days over the latest 30-year period (1986–2016). Furthermore, the interrelationship between various phenological stages and floral morphogenesis of the same cultivars in the years 2013 and 2014 was studied, and related to flower-bud formation. The different stages of floral morphogenesis were identified, presented and used as a basis for assessment of flower-bud development in serial dissections. Floral initiation in spurs of actively growing trees took place in late July, approx. 8 weeks after full bloom, whereas in extension shoots, initiation took place after growth cessation in August, approximately 2 weeks later. Decreasing temperatures in mid-August coincided with the cessation of growth and floral initiation in extension shoots, suggesting that the processes may be causally related. The results show that, in the Nordic climate, the period between blooming and initiation of new floral primordia is considerably shorter than in warmer climates, suggesting that the period is only marginally long for completion of shoot growth, and hence, timely floral initiation. This may be an important reason for the irregular flowering and frequent incidences of alternate bearing in apple trees in the cool Nordic climate. The presented information can be useful for determination of the correct timing of flower and fruit thinning to counterbalance biennial bearing.     

Evaluation of the discriminance capacity of RAPD, isoenzymes and morphologic markers in apple (Malus x domestica Borkh.) and the congruence among classifications

Twenty-one apple accessions were characterised and classified taxonomically with randomly amplified polymorphic DNA (RAPD). The results were compared with morphological and isoenzymatic classifications of the same material by calculating the congruence between similarity matrices. The results confirmed the greater discrimination capacity of RAPD compared with isoenzymes, although the taxonomic clusters obtained with both types of markers were congruent. The genetic relationships calculated using RAPD and isoenzymes showed little congruence with the morphological relationships among accessions. Isoenzymatic markers are useful in germplasm banks to evaluate the diversity of newly introduced accessions. The use of RAPD markers are especially beneficial to discriminate between material that is genetically similar, to evaluate genetic variability within a collection and to choose the components of the core collection.     

Impact of suppression of ethylene action or biosynthesis on flavor metabolites in apple (Malus domestica Borkh) fruits

To understand the role of ethylene in regulating the overall flavor of apple fruits, ethylene production or action was reduced using transgenic apple trees suppressed for ACC-synthase or ACC-oxidase enzyme activity or by the addition of 1-methylcyclopropene (1-MCP), an ethylene action inhibitor. Flavor components were differentially regulated in response to the suppression of both ethylene biosynthesis and action. Headspace analysis of aroma production, an ethylene-associated event, showed a reduction in ester and alcohol production in the ethylene-suppressed lines and in the apples treated with 1 microL L(-1) 1-MCP for 20 h at 20 degrees C. However, no major differences were observed in the concentrations of aldehyde volatiles. Other flavor metabolites that showed an ethylene-dependent pattern were organic acids and sugars. Malic acid degradation was significantly reduced under ethylene-suppressed conditions, showing a recovery after the fruit was exposed to ethylene. Sucrose and fructose concentrations were influenced by suppression or enhancement of ethylene. Total phenolics as well as individual phenolic compounds showed an ethylene-dependent regulation only in response to the suppression of ethylene biosynthesis, but not when ethylene action was inhibited.     

Free galactose concentrations in fresh and stored apples (Malus domestica) and processed apple products

Gas chromatography was used to quantitate free galactose in Braeburn, Fuji, Red Delicious, and Spartan apples during cold storage, after thermal processing of apple slices and in juice produced using clarification and/or liquifaction enzymes. Spartan had significantly higher galactose levels as compared to Red Delicious apples, but changes in galactose in all varieties during 9 months of cold storage were insignificant. Blanching and canning decreased galactose levels, but doubling the thermal processing during canning increased the free galactose concentration detected in plant tissue. An enzymatic liquefaction aid used to prepare apple juice dramatically increased the free galactose content while a clarification aid caused only a slight increase due to its selective action on soluble pectin. These findings provide useful information for dietitians to base diet recommendations for galactosemic patients.     

苹果SUPERMAN类蛋白家族的基因克隆与生物信息学分析

SUPERMAN 类基因是植物生长发育中的重要转录因子。为了揭示苹果 SUPERMAN 类蛋白家族的生理基因功能,本研究以富士苹果(Malus domestica Borkh.cv. Fuji)为试材,利用 RT-PCR 方法和锚定PCR 方法克隆了 MdLIFa(GenBank 登录号 HM370493)和启动子序列,利用特异引物 PCR 方法克隆了苹果基因组中其余 11 个 SUPERMAN 类基因。12 个基因命名为 MdLIFa/MdLIFb (GenBank 登录号:HQ260429)、MdSUP1a (HQ418477)/MdSUP1b (HQ418478)、MdSUP4 (HQ418475)/MdSUP12 (HQ418476)、MdSUP5a (HQ418469)/MdSUP13b (HQ418470)、MdSUP9a (HQ418473)/MdSUP9b (HQ418474) 和 MdSUP5b(HQ418471)/MdSUP13a(HQ418472),分别位于 11/3、1/1、4/12、5/13、9/ 未知和 5/13 号染色体,每对基因间的同源性均在86%以上。通过基因枪法轰击洋葱(Allium cepa)表皮细胞,基因瞬时表达证明 MdLIFa 定位于细胞核内。MdLIFa 基因启动子序列中含有多个参与花粉发育、根特异性表达、与光诱导和 Dof 单锌指基因家族作用的调控元件等。系统进化分析表明,MdLIFa/b、MdSUP9a/9b 和 MdSUP5a/13b、以及MdSUP1a/b、MdSUP5b/MdSUP13a和 MdSUP4/12 分别聚类为一组,参与植株的形态建成和花器官发育。苹果 12 个 SUPERMAN 类基因成对高度同源,具有生物学功能的多样性,为深入的基因鉴定和遗传转化提供有用信息。     

Impacts of farmland conversion to apple (Malus domestica) orchard on soil organic carbon stocks and enzyme activities in a semiarid loess region

Land‐use change often affects the sizes of soil organic carbon (SOC) stocks and the activities of soil enzymes. Responses of relevant soil quality indices caused by farmland conversion to orchard are largely unknown in the semiarid loess regions. This study was conducted at orchard sites, which have been under very intensive cultivation, to evaluate the impacts of farmland conversion to apple (Malus domestica) orchard on SOC stocks and soil enzyme activities in the semiarid loess region of Weibei, Shaanxi province, China. The spatial and temporal changes in a variety of soil quality indices were measured for the 0–100 cm soil profile in apple orchards of three age groups (< 10 y, juvenile; 10–15 y, mature; > 15 y, over‐mature) and adjacent farmlands (control). After farmland conversion, total SOC (TOC) content and density and soil alkaline phosphatase activity significantly decreased, while soil catalase activity increased for the 0–100 cm soil profile. The labile SOC (LOC) content, its proportion to TOC content, and carbon management index (CMI, changes in the total content and lability of SOC) significantly increased in the 0–40 cm soil layer, whereas soil urease and invertase activities only increased in the 0–20 cm layer (P < 0.05). With increasing age of apple orchards, SOC stocks significantly increased after 10 y, being more than 10% larger in mature and over mature orchards than in adjacent farmlands. The LOC content and CMI value also had an increasing trend, while soil enzyme activities showed different response patterns. There were significant correlations between soil enzyme activities, SOC fractions, and CMI value (P < 0.05). We concluded that farmland conversion to apple orchard affected soil quality by reducing SOC stocks in the soil profile and changing SOC content as well as soil enzyme activities at various depth intervals. Long‐term apple cultivation was effective to restore SOC stocks, although it took over a decade to rebuild a new increasing trend after farmland conversion.     

钙对苹果果实超氧化物歧化酶、过氧化氢酶活性及其基因表达的影响

采用果实薄片培养试验研究不同钙浓度(0、1和10 mmo1/L CaCl2 ;5 mmo1/L EGTA)和处理时间(6、12、24和48 h)下苹果果实活性氧代谢特征;运用荧光定量PCR方法分析苹果超氧化物歧化酶（SOD）、过氧化氢酶（CAT）基因在分子水平的表达。高钙处理12 h后,SOD酶的活性显著增加,O2.-产生速率显著下降;高钙处理24 h后,CAT酶的活性显著增加,H2O2含量显著下降;缺钙处理下SOD酶和CAT酶的活性受到抑制,O2.-形成速率和H2O2含量显著增加。基因表达实验显示,高钙处理12h后,SOD基因的表达量增加,与SOD酶活性变化一致,说明SOD酶的活性取决于SOD基因的表达量;高钙处理12 h后,CAT1基因的表达量增加,而CAT酶的活性是在加钙处理24 h后显著增加,说明CAT酶的活性并不完全取决于CAT1基因的表达量,推断其酶活性还取决于该基因翻译后的修饰调控。上述研究表明苹果外源补钙通过在分子水平上调SOD和CAT1基因表达量来激活SOD和CAT酶的活性,有效减少体内活性氧积累,确保果实生理代谢平衡。     

Chemical composition of volatile extract and biological activities of volatile and less-volatile extracts of juniper berry (Juniperus drupacea L.) fruit

Volatile chemicals in a dichloromethane extract from a steam distillate of juniper berry fruit (Juniperus drupacea L.) and its two column chromatographic fractions (eluted with hexane and ethyl ether) were analyzed by gas chromatography/mass spectrometry. The major compounds in the dichloromethane extract were alpha-pinene (23.73%), thymol methyl ether (17.32%), and camphor (10.12%). A fraction eluted with hexane contained alpha-pinene (44.24%) as the major constituent. A fraction eluted with ethyl ether had thymol methyl ether (22.27%) and camphor (19.65%) as the main components. Three samples prepared from the distillate and two additional samples prepared by petroleum ether and ethanol extraction directly from juniper berry fruits exhibited clear antioxidant activities with dose response in both 1,2-diphenyl picrylhydrazyl and beta-carotene assays. All samples except the hexane fraction showed comparable activities to that of the synthetic antioxidant t-butyl hydroquinone at a level of 200 microg/mL in the two testing systems. The extracts of dichloromethane, petroleum ether, and ethanol exhibited appreciable antimicrobial activities against six microorganisms with minimum inhibitory concentrations ranging from 0.5 mg/mL (volatile extract against Candida albicans ) to 1.2 mg/mL (ethanol extract against Aspergillus niger ). The results of the present study suggest that this fruit could be a natural antioxidant supplement for foods and beverages.     

Morpho-physio and anatomical characterization of the indigenous Himalayan crabapple: Malus baccata (L.) Borkh. (Rohru) and exotic Malus pumila Mill. for dwarfing traits

Genetic Resources and Crop Evolution - The wild Malus germplasm is considered as a gene reservoir for various biotic and abiotic stresses tolerance/resistance genes, including important novel...     

Phenolic profile, antioxidant property, and anti-influenza viral activity of Chinese quince (Pseudocydonia sinensis Schneid.), quince (Cydonia oblonga Mill.), and apple (Malus domestica Mill.) fruits

To evaluate the phenolic extracts of Chinese quince, quince, and apple fruits, their phenolic profiles, antioxidant properties, and anti-influenza viral activities were investigated. Chinese quince had the largest amount of phenolics consisting mainly of high polymeric procyanidins. Quince had considerable amounts of hydroxycinnamic derivatives mainly composed of 3-caffeoylquinic acid and 5-caffeoylquinic acid and polymeric procyanidins. Apple (cv. Fuji) had the lowest amount of phenolics, mainly 5-caffeoylquinic acid and monomeric and oligomeric procyanidins. The antioxidant functions of Chinese quince and quince phenolic extracts were superior to that of chlorogenic acid standard or ascorbic acid evaluated in both the linoleic acid peroxidation system and the DPPH radical scavenging system. However, those extracts were less effective than apple phenolics or (-)-epicatechin in linoleic acid peroxidation system. On the other hand, Chinese quince phenolics showed the strongest anti-influenza viral activity on the hemagglutination inhibition test.     

Diversity of the European indigenous wild apple (Malus sylvestris (L.) Mill.) in the East Ore Mountains (Osterzgebirge), Germany: II. Genetic characterization

In the present study genetic diversity and hybridization with cultivars were investigated in a population of the endangered European wild apple species Malus sylvestris (L.) Mill. with the aim to establish a basis for the implementation of conservation activities and to ensure its long-term preservation. A total of 284 putative M. sylvestris trees located in the East Ore Mountains were investigated along with a standard set of reference apple genotypes proposed by the European Cooperative Program for Plant Genetic Resources (ECPGR) and 13 old apple cultivars often cultivated in Saxony. The genetic analysis was performed using 12 microsatellite markers also recommend by the ECPGR. To differentiate ‘true type’ M. sylvestris individuals, hybrids and apple cultivars (Malus × domestica Borkh.) a model-based cluster analysis was performed using STRUCTURE. Two clusters were identified consisting of M. sylvestris and M. × domestica genotypes. About 40 % of the putative M. sylvestris showed an admixture of the species-specific allele frequencies and were defined as hybrids. The genetic diversity of the ‘true type’ M. sylvestris population was still high but slightly lower than in the apple cultivars especially since some SSR loci were fixed on one or few alleles in the M. sylvestris population. The differentiation parameters between ‘true type’ wild apple and cultivars indicated a clear discrimination between the wild and cultivated apple individuals. This fact confirms our expectation of the existence of ‘true type’ M. sylvestris individuals in the East Ore Mountains and argues for the realization of preservation measures in this area.     

Release of nitrogen,phosphorus, and potassium during the decomposition of apple (Malus domestica) leaf litter under different fertilization regimes in Loess Plateau,China

Hydrofluoric acid (HF) is one of the air-pollutants known to be harmful to plants, and is emitted by industries and volcanoes. Fluoride affects plant metabolism and its effects on respiration are fully understood (2). However, little is known about the effect of fluoride on photosynthesis as has been reviewed by Chang (2). Fluoride reduces syntheses of chlorophyll and protochlorophyll, induces the degradation of chloroplast structure in bush bean and soybean (5), and inhibits the Hill reaction (1). When orange leaves were fumigated with HF, the main site of fluoride accumulation was the chloroplasts (3). Although it may therefore be presumed that the rate of photosynthesis of intact leaves decreases during HF fumigation, fluoride-effects on APS have not been fully determined.