Peroxidase activity and superficial scald development in apple fruit

The relationship between soluble peroxidase (EC 1.11.1.7; POX) activity and the development of a chilling-related disorder, superficial scald, was studied in three apple fruit (Malus x domestica Borkh.) systems: a White Angel x Rome Beauty population with progeny with different scald susceptibilities; Delicious from three harvests with progressively declining scald susceptibility; and the scald-resistant Idared and the scald-susceptible Law Rome. Differences in incidence and severity of scald in progeny from White Angel x Rome Beauty progeny tended to show relationships with POX activity at harvest, but, overall, associations were not consistent. However, greater scald incidence and lower POX activity were found in less mature Delicious fruit than in later harvested fruit. Also, the scald-resistant Iotadared had a much higher POX activity compared with the scald-susceptible Law Rome. A general hypothesis that POX activity is related to scald susceptibility was generally supported, but exceptions were observed.     

Effect of superficial scald suppression by diphenylamine application on volatile evolution by stored Cortland apple fruit

Cortland apple fruit (Malus x domestica Borkh.) stored for 120-140 days in air at 0 degrees C were warmed to 22 degrees C and held for 8 days. A portion of the fruit was dipped in a solution of diphenylamine (DPA) at harvest to prevent scald development. Scald occurred only in those fruit not treated with DPA, and its development was accelerated after transfer to 22 degrees C. Ester production from apple fruit tended to increase from day 0 to day 6 of poststorage holding and declined thereafter in both treatments. However, ester production in scald-developing fruit was reduced by approximately 50%. The reduction in volatile production remained relatively constant during the rapid development of scald symptoms. Furthermore, the reduction in volatile production appeared to be independent of respiration and ethylene production. Production of esters derived from hexanol was most reduced in fruit developing scald, with hexyl 2-methylbutanoate production being reduced approximately 15-fold. Interestingly, the production of methyl butanoate was detected only in scalding fruit. alpha-Farnesene production in fruit developing scald was reduced 43% compared with DPA-treated fruit. In contrast, the primary volatile oxidation product of alpha-farnesene, 6-methyl-5-hepten-2-one (MHO), was present only in fruit developing scald. The data suggest that inhibition of ester production may occur as a result of the physiological changes associated with susceptibility to, rather than expression of, scald symptoms.     

Relationship of superficial scald development and alpha-farnesene oxidation to reactions of diphenylamine and diphenylamine derivatives in Cv. Granny Smith apple peel

Cv. Granny Smith apple fruit, treated at harvest with aqueous emulsions containing diphenylamine (DPA) and DPA derivatives, were evaluated for the peel disorder superficial scald (scald) after 6 months of cold storage at 1 degrees C plus 0 or 7 days at 20 degrees C. Metabolism of these derivatives and alpha-farnesene oxidation were also evaluated after 6 months. Derivatives substituted at the para position prevented scald, but scald developed on fruit treated with derivatives substituted in the amino, ortho, or meta positions. The extent of scald control was also dependent on the chemical nature of the functional group used to derivatize DPA. Hydroxylation of DPA and DPA derivatives during storage was not associated with scald control. Methoxylated DPA derivatives produced during storage resulted from O-methylation of C-hydroxylated derivatives rather than C-methoxylation of DPA. N-Nitrosodiphenylamine provided partial scald control, possibly resulting from its degradation to DPA, indicating that the amino hydrogen of DPA may be crucial for scald control. Results suggest that functional group position and chemical properties both contribute to the efficacy of DPA derivatives for scald control.     

Conjugated triene oxidation products of alpha-farnesene induce symptoms of superficial scald on stored apples.

The sesquiterpene alpha-farnesene (1) and its conjugated triene oxidation products accumulate in the skin of apples after harvest and are implicated as the causal agents of superficial scald. Conjugated triene oxidation products and analogues were synthesized and applied to the surface of Granny Smith apples either as vapors or in squalane. Farnesyl hydroperoxide (2a), trienol (2b), endoperoxide (3a), dehydronerolidol (5), and cumyl hydroperoxide (4) all produced the symptoms of superficial scald when applied at nanomolar doses. Scald-inducing activity was dependent on the mode of application. Farnesyl hydroperoxide (2a) was the most active conjugated triene when applied directly to the apple skin. Trienol (2b) also induced scald symptoms and partially reversed the inhibition of scald caused by diphenylamine.     

贮前苯胺蓝染色预测苹果冷藏末期表皮病害

为了实现对苹果贮藏末期表皮病害的预测,以红富士苹果（Malus domestica Borkh cv. Fuji）为试材,通过苯胺兰染色的方法,研究了苹果采后苯胺兰染色与果实贮藏末期几种表皮病害的关系。结果表明：富士苹果采后苯胺兰染色指数与贮藏末期的好果率呈明显负相关,相关系数为-0.947;果实苯胺兰染色的级别与二氧化碳伤害指数和腐烂指数密切相关,其决定系数分别为0.9779和0.8394,但苯胺兰染色的级别与贮藏末期果实的虎皮病和皮孔斑点病之间无明显的相关性。因此,苹果采后苯胺兰染色可用于预测果实贮藏末期的好果率。     

Temperature-dependent autoxidation of conjugated trienols from apple peel yields 6-methyl-5-hepten-2-one, a volatile implicated in induction of scald

Conjugated triene (CT) oxidation products of alpha-farnesene have long been thought to be involved in development of superficial scald in apple fruit. Early studies found that CT hydroperoxides and the volatile 6-methyl-5-hepten-2-one (MHO) are major in vitro autoxidation products of alpha-farnesene. However, it was recently shown that > or =99% of the oxidation products of alpha-farnesene that accumulate in apple peel are conjugated trienols (CTols), isomers of 2,6,10-trimethyldodeca-2,7,9,11-tetraene-6-ol. HPLC-purified CTols from fruit of two scald-susceptible cultivars, Granny Smith (GS) and Red Delicious (RD), were used to study autoxidation of these compounds in vitro. Incubation of CTols in sealed glass vials under air resulted in accumulation of MHO. Oxygen enrichment did not increase the amount of MHO produced. Regardless of which cultivar CTols were derived from, at 0 degrees C autoxidation yielding MHO was quite slow and linear, whereas at 20 degrees C MHO production was much more rapid, and after several hours the rate increased abruptly. However, CTols isolated from GS and RD fruit differed in the duration of the initial lag phase and the overall level of MHO generated at 20 degrees C. The sharp increase in MHO production occurred after 3 h with GS CTols and at about 12 h with RD CTols. Also, the yield of MHO from GS CTols after 6 h at 20 degrees C was nearly 6-fold greater than that from RD CTols after 20 h at 20 degrees C. The antioxidants butylated hydroxytoluene and diphenyamine reduced the yield of MHO by about 97%. Recent work has shown that MHO can induce scald-like symptoms in apple peel and that tissue sensitivity increases with time in storage. This may explain the correlation between high CTol levels and scald development, and why symptoms rapidly intensify when fruits are removed from cold storage.     

Cloudy apple juice is more effective than apple polyphenols and an apple juice derived cloud fraction in a rat model of colon carcinogenesis

As recently shown, a cloudy apple juice (CloA) was effective to modulate colon cancer associated parameters in rats treated with 1,2-dimethylhydrazine (DMH). To identify the bioactive substance classes in CloA, we fractionated CloA to yield a total polyphenol (PF) and a cloud (CF) fraction consisting of proteins, fatty acids, polyphenols, and cell wall polysaccharides. Rats received water (control (Cont)) or CloA, PF, and CF separate or combined (PF-CF) ad libitum for 7 weeks starting one week before the first DMH-injection. As determined by comet assay, the DMH-induced genotoxicity in colonocytes of controls (Cont/DMH: 7.7 +/- 0.5%) was significantly reduced by CloA (3.3 +/- 0.3%) but not by any of the fractions. The crypt cell proliferation induced by DMH (Cont/NaCl: 7.5 +/- 0.6%; Cont/DMH: 14.9 +/- 0.8%) was significantly decreased by CloA (9.4 +/- 0.4%), PF (12.4 +/- 0.7%), CF (11.6 +/- 0.4%), and PF-CF (12.4 +/- 0.6%). Although not statistically significant, CloA tended to reduce the number of large aberrant crypt foci (ACF) (Cont/DMH: 19.0 +/- 3.7; CloA/DMH: 12.3 +/- 1.9), while none of the fractions affected ACFs. Neither CloA nor the fractions changed mRNAs of colonic cyclooxygenases (COX-1, COX-2), glutathione-associated enzymes (GST-M2, gamma-GCS, GST-P), the splenocyte CD4/CD8 ratio, natural killer cell activity, and plasma antioxidant status. These results demonstrate that CloA had a higher cancer-preventive potential than the fractions and further, besides PF, identified CF as an additional bioactive fraction of CloA.     

Antioxidant properties of green tea extract protect reduced fat soft cheese against oxidation induced by light exposure

The effect of two different antioxidants, EDTA and green tea extract (GTE), used individually or in combination, on the light-induced oxidation of reduced fat soft cheeses (0.2 and 6% fat) was investigated. In samples with 0.2% fat, lipid hydroperoxides as primary lipid oxidation products were not detected, but their interference was suggested from the formation of secondary lipid oxidation products such as hexanal and heptanal. The occurrence of these oxidation markers was inhibited by spiking with 50 ppm EDTA or 750 ppm GTE, or a combination of the two prior to irradiation. In contrast, addition of 50 ppm EDTA to samples with 6% fat was ineffective, but 750 ppm GTE (alone or in combination with EDTA) strongly reduced levels of hexanal and heptanal. Accumulation of primary lipid hydroperoxides was not affected by GTE, hence antioxidative activity was ascribed to scavenging of hexanal and heptanal precursors. These radical intermediates result from hydroperoxide disintegration, and subsequent scavenging by GTE, which acts as a radical sink, corroborates the intense signal observed by electron paramagnetic resonance (EPR) spectroscopy.     

A multi-phasic approach reveals that apple replant disease is caused by multiple biological agents, with some agents acting synergistically

Apple replant disease (ARD) has been reported from all major fruit-growing regions of the world, and is often caused by a consortium of biological agents. The aim of this study was to investigate the etiology of ARD in South Africa in six orchard soils, using a multiphasic approach under glasshouse conditions. This approach first involved determining the ARD status of the soils by monitoring apple seedling growth responses in non-treated soil versus growth in pasteurized soil, as well as in 15% non-treated soil that was diluted into pasteurized soil. Subsequently, the potential for specific organisms to function as causal agents of ARD was investigated using (i) biocide applications, (ii) quantitative real-time PCR (qPCR) analyses of ARD ‘marker’ microbes (Pythium irregulare, Pythium sylvaticum, Pythium ultimum, Pythium vexans, Rhizoctonia solani AG-5 and the genera Cylindrocarpon and Phytophthora), (iii) nematode analyses, (iv) isolation of actinomycetes and (v) pathogenicity testing of actinomycetes individually, and when co-inoculated with P. irregulare or Cylindrocarpon macrodidymum. The analyses showed that the soils could be grouped into low, moderate and severe ARD soils, with each group containing two soils. Several lines of evidence suggested that actinomycetes are not involved in ARD in South Africa. Multiple biological agents were determined to contribute to ARD including (i) oomycetes (Phytophthora and Pythium) that are important based upon their widespread occurrence, and the fact that metalaxyl application improved seedling growth in four soils (ii) the genus Cylindrocarpon that was also widespread, and for which a synergistic interaction with P. irregulare was demonstrated and (iii) occasionally parasitic nematodes, mainly Pratylenchus penetrans, Pratylenchus scribneri and Pratylenchus delattrei, since fenamiphos application improved seedling growth in two orchards. qPCR analyses of the ARD marker microbes showed that R. solani AG-5 is absent from South African orchards, and that P. ultimum is widespread, even though the latter species could not be detected in previous isolation studies. The other marker microbes were also widespread, with the exception of P. sylvaticum. qPCR quantification of the marker microbes could not be correlated with the severity of ARD in any manner. qPCR analyses did, however, show that possible root pruning pathogens such as P. irregulare, P. sylvaticum and P. ultimum had much lower DNA concentrations in seedling roots than P. vexans and the genera Cylindrocarpon and Phytophthora.     

苹果腐心病的透射光谱在线检测系统设计及试验

针对苹果内部缺陷在线检测的产业技术需求,研究基于透射光谱技术的苹果内部缺陷在线检测系统。研究设计了光源套件、专用光纤和果托式输送单元等关键部件,提升在线透射光谱质量、降低热损伤和机械损伤;解决了光电信号干扰问题,开发了专用检测软件,实现苹果内部品质信息的无损在线获取。比较分析了正常苹果与腐心病果的光谱响应差异,优化参数后设置在线检测速度3个/秒,触发控制光谱采集时间80 ms。在选择特征波长的基础上利用线性判别分析建立了苹果腐心病的在线判别模型,预测的总体识别率达90%以上。研究结果表明该系统可以实现苹果内部缺陷的快速、无损在线检测。     

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.     

Carbon mineralization is promoted by phosphorus and reduced by nitrogen addition in the organic horizon of northern hardwood forests

Limitations to the respiratory activity of heterotrophic soil microorganisms exert important controls of CO₂ efflux from soils. In the northeastern US, ecosystem nutrient status varies across the landscape and changes with forest succession following disturbance, likely impacting soil microbial processes regulating the transformation and emission of carbon (C). We tested whether nitrogen (N) or phosphorus (P) limit the mineralization of soil organic C (SOC) or that of added C sources in the Oe horizon of successional and mature northern hardwood forests in three locations in central New Hampshire, USA. Added N reduced mineralization of C from SOC and from added leaf litter and cellulose. Added P did not affect mineralization from SOC; however, it did enhance mineralization of litter- and cellulose- C in organic horizons from all forest locations. Added N increased microbial biomass N and K₂SO₄-extractable DON pools, but added P had no effect. Microbial biomass C increased with litter addition but did not respond to either nutrient. The direction of responses to added nutrients was consistent among sites and between forest ages. We conclude that in these organic horizons limitation by N promotes mineralization of C from SOC, whereas limitation by P constrains mineralization of C from new organic inputs. We also suggest that N suppresses respiration in these organic horizons either by relieving the N limitation of microbial biomass synthesis, or by slowing turnover of C through the microbial pool; concurrent measures of microbial growth and turnover are needed to resolve this question.     

Surface functional properties of native, acid-treated, and reduced soy glycinin. 2. Emulsifying properties

Emulsifying properties of native and chemically modified soy glycinins were studied. The influence of ionic strength, protein sample composition and concentration, and assay conditions on the flocculation-creaming process and coalescence resistance was analyzed. Differences in these emulsifying properties were exhibited by native glycinins, which have a variable content of 4S, 11S, and 15S forms. Structure and functionality of native glycinin were modified by means of combined treatments: mild acidic treatments without heating or with heating at variable time and with or without disulfide bonds reduction. Modified glycinins presented different degrees of deamidation, surface hydrophobicity, and molecular mass. A slight enhancement of emulsifying stability at moderated deamidation degrees was observed. In different protein samples, a positive relationship between the flocculation-creaming rate constant and equilibrium oil volume fraction of emulsions with surface hydrophobicity was detected. A remarkable difference was observed between reduced and nonreduced samples, mainly with respect to behavior at low or high ionic strength.     

Surface functional properties of native, acid-treated, and reduced soy glycinin. 1. Foaming properties

Foaming properties of native and chemically modified glycinin were evaluated. Effects of ionic strength and glycinin composition and concentration on foam formation and stabilization were studied. Glycinin was modified by means of combined treatments: cold or hot acidic treatments, with or without later disulfide bridges reduction. Modified proteins obtained from glycinin present different degrees of dissociation, deamidation, and as consequence, varied surface hydrophobicity and molecular size. Parameters of forming and stabilizing of foam were correlated with both deamidation and dissociation degrees of modified and native glycinin samples. A positive relationship was observed between surface behavior and foaming properties of different protein species. Results show that dissociation, deamidation, and reduction have produced structural changes on glycinin (increased surface hydrophobicity, increased net charge, decreased molecular size) which enhance the adsorption and anchorage of proteins at the air-water interface and, consequently, improve the foam forming and stabilizing capacities.     

Nutrient status of apple orchards in Canterbury,New Zealand. 2. Specific problems

Abstract

In a survey of the nutrient status of Cox's Orange Pippin and Braeburn apple orchards in the Canterbury region of New Zealand three separate problems were identified in a number of orchards. These were low soil pH in the herbicide strip area, low concentrations of fruit Ca and low concentrations of leaf Zn.

Seven out of 26 Cox's Orchard sites had a significantly lower soil pH (0–10 cm) in the herbicide strip than the grassed alley. At some sites soil acidification had occurred down to a depth of 30 cm. The major cause of such acidification was thought to be nitrification of ammonium originating from both fertilisers and mineralisation of soil organic matter.

No one reason was responsible for low concentrations of Ca (< 2.5 mg/100 g) in cortical plugs of apple tissue. At one site it was related to a deficiency of soil Ca but at two other sites, levels of exchangeable Ca were relatively high. Levels of exchangeable Mg and/or K were however exceptionally high suggesting that a cation imbalance in the soil may have contributed to the low fruit Ca. At other sites, low fruit Ca was probably related to factors other than soil nutrient status such as fruit size and possibly water stress during summer.

In some cases, low concentrations of leaf Zn were associated with low extractable soil Zn coupled with relatively high levels of soil P. One orchard had a reasonably high level of extractable soil Zn but an exceptionally high level of extractable soil P probably inhibited Zn uptake by the tree. At another site lime‐induced Zn deficiency was evident whilst at another, exceptionally high levels of extractable soil Cu may have inhibited Zn accumulation by trees.     

Isolation, characterization, and antioxidant activity of E- and Z-p-coumaryl fatty acid esters from cv. Annurca apple fruits

A total of 12 fatty acid esters of Z- and E-p-coumaryl alcohol were isolated from cv. Annurca apple fruit and characterized. This apple variety is widely cultivated in the south of Italy, and the fruits typically undergoe a reddening treatment after harvest. Structures of the p-coumaryl esters were elucidated by GC-MS and (1)H and (13)C NMR after purification of individual compounds by HPLC. It was found that the esters are localized in the fruit peel. During reddening of the fruit, there was a substantial increase in the amount of esters and particularly in molecular species with unsaturated fatty acids. The individual compounds were tested for antioxidant activity, and over half were shown to be at least as effective as alpha-tocopherol.     

Study of the phenolic profile of cider apple cultivars at maturity by multivariate techniques.

Phenolic compounds in 46 Spanish cider apple varieties were determined by RP-HPLC with direct injection. Several pattern recognition procedures, including principal component analysis (PCA), linear discriminant analysis (LDA), and partial least squares (PLS-1), were applied to the data in an attempt to classify the samples into bitter and nonbitter categories. Reliable decision rules were obtained by both LDA and PLS-1. LDA achieved 91.3 and 85.7% correct classification respectively, for internal and external evaluation of the model.