Allelic polymorphism and inheritance of MdACS1 and MdACO1 genes in apple (Malus × domestica Borkh.)

The allelic polymorphism and inheritance of MdACS1 and MdACO1 genes were analysed in 28 apple cultivars, which were derived from reciprocal crosses of the following parental pairs: ‘Golden Delicious’ × ‘James Grieve’, ‘Golden Delicious’ × ‘Jonathan’, ‘Cox's Orange Pippin’ × ‘Golden Delicious’, ‘Cox's Orange Pippin’ × ‘Jonathan’. Polymorphisms were detected by PCR and the use of two restriction enzymes (BamH1 and RsaI). In addition, new primers were designed for the further discrimination of MdACO1 alleles. Two alleles of MdACS1 gene (MdACS1‐1 and MdACS1‐2) and three alleles of MdACO1 gene (a, b and c) were detected. Cloning and sequencing of MdACO1 alleles confirmed a high conservation and some differences within the coding regions and helped to reconcile between different numbering systems. Observed segregations confirmed that alleles a, b and c belonged to the MdACO1 gene. It is apparent that polymorphisms within the MdACS1 and MdACO1 genes could aid cultivar genotyping and identification and, furthermore, that the MdACS1‐2/2 genotype is correlated with a long apple storage life.     

Influence of time from harvest to 1-MCP treatment on apple fruit quality and expression of genes for ethylene biosynthesis enzymes and ethylene receptors

A strong potent inhibitor of ethylene action, 1-methylcyclopropene (1-MCP) maintains apple fruit quality during storage. To understand the influence of time after harvest until 1-MCP treatment, we studied expression patterns of genes for ethylene biosynthesis enzymes and ethylene receptors in two apple cultivars, ‘Orin’ and ‘Fuji’, which differ in ethylene production. Ethylene production and expression of MdACS1, MdERS1, and MdERS2 were suppressed in all 1-MCP-treated ‘Fuji’ fruit, but in ‘Orin’, the later 1-MCP was applied after harvest, the less was the suppression of ethylene production and expression of these genes. In fruit in which 1-MCP had low efficacy (e.g., ‘Orin’ treated at 7 DAH), ethylene production and the level of MdERS1 were briefly reduced by 1-MCP treatment at 2 days after treatment, then began to increase. Since ethylene receptors negatively regulate the ethylene signalling pathway, the increased levels of ethylene production and ethylene receptors after 1-MCP treatment might reduce 1-MCP efficacy.     

Effects of preharvest nitric oxide treatment on ethylene biosynthesis and soluble sugars metabolism in ‘Golden Delicious’ apples

In order to examine the influence of preharvest nitric oxide (NO) treatment on ethylene biosynthesis and soluble sugar metabolism in ‘Golden Delicious’ apples, apple trees were sprayed with 50 μM sodium nitroprusside (SNP) (a donor of NO) 14 days before harvest. The results indicated that preharvest SNP treatment can increase the NO content and the NOS activity in apple fruit, therefore, delay the accumulation of ethylene due to its inhibition on the activities of 1-aminocyclopropane-1-carboxylate synthase (ACS) and 1-aminocyclopropane-1-carboxylate oxydase (ACO). Fructose is the main sugar in ‘Golden Delicious’ apple. The synthesis of sucrose was stimulated and the decomposition of sucrose was inhibited by this treatment, thus causing the accumulation of sucrose. We can draw a conclusion that pre-harvest SNP (50 μM) treatment can increase the NO content of fruit during storage, while higher NO content can further regulate fruit ripening through its effect on ethylene and sugar metabolism in ‘Golden Delicious’ apple fruit during storage at 18 °C.     

Efficacy of 1-MCP treatment in tomato fruit: 2. Effect of cultivar and ripening stage at harvest

Four cultivars of tomato fruit (‘Cherry’, ‘Daniela’, ‘Patrona’ and ‘Raf’) were harvested at two ripening stages (S1 and S2), treated with 0.5 μl l⁻¹ of 1-methylcyclopropene (1-MCP) for 24 h and stored at 10 °C for 28 days. For all cultivars, control fruit deteriorated very rapidly (due to weight loss, softening, colour changes and decay) with an estimated shelf life of 7 days (‘Cherry’ and ‘Patrona’) and 14 days (‘Daniela’ and ‘Raf’), independently of the ripening stage at harvest. All quality parameters for all cultivars were delayed and/or inhibited in treated fruit, the efficacy of 1-MCP being higher in tomatoes harvested at the S2 ripening stage. At this stage, the organoleptic properties had already developed in fruit on the plant and tomatoes could thus reach consumers with optimal postharvest quality.     

Characterization of ethylene biosynthesis and its regulation during fruit ripening in kiwifruit,Actinidia chinensis ‘Sanuki Gold’

Ethylene biosynthesis in kiwifruit, Actinidia chinensis ‘Sanuki Gold’ was characterized using propylene, an ethylene analog, and 1-methylcyclopropene (1-MCP), an inhibitor of ethylene perception. In fruit harvested between a young stage (66 days after pollination) (DAP) and an early commercial harvesting stage (143 DAP), 2 days of exposure to propylene were sufficient to initiate ethylene biosynthesis while in fruit harvested at commercial harvesting stage (154 DAP), 4 days of propylene treatment were required. This observation suggests that response of ethylene biosynthesis to propylene treatment in kiwifruit declined with fruit maturity. Propylene treatment resulted in up-regulated expression of AC-ACO1, AC-ACO2, AC-SAM1 and AC-SAM2, prior to the induction of AC-ACS1 and ethylene production, confirming that AC-ACS1 is the rate limiting step in ethylene biosynthesis in kiwifruit. Treatment of fruit with more than 5 μL L^?1 of 1-MCP after the induction of ethylene production subsequently suppressed ethylene production and expression of ethylene biosynthesis genes. Treatment of fruit with 1-MCP at harvest followed with propylene treatment delayed the induction of ethylene production and AC-ACS1 expression for 5 days. These observations suggest that in ripening kiwifruit, ethylene biosynthesis is regulated by positive feedback mechanism and that 1-MCP treatment at harvest effectively delays ethylene production by 5 days.     

1-MCP regulates ethylene biosynthesis and fruit softening during ripening of ‘Tegan Blue’ plum

To investigate the effects of postharvest application of 1-MCP on ethylene production and fruit softening, activities of ethylene biosynthesis and fruit softening enzymes were measured during postharvest ripening of plum (Prunus salicina Lindl. cv. Tegan Blue) fruit after being exposed to 1-MCP (0, 0.5, 1.0 or 2.0 μL L⁻¹) at 20 ± 1 °C for 24 h. Following the treatments, fruit were allowed to ripen at ambient temperature (20 ± 1 °C), and ethylene production in fruit, activities of ACS and ACO, ACC content and fruit softening enzymes (PE, EGase, exo-PG and endo-PG) in fruit skin and pulp were recorded at different intervals. Postharvest application of 1-MCP significantly delayed and suppressed the climacteric ethylene production with reduction in the activities of ethylene biosynthesis enzymes (ACS, ACO) and ACC content, and fruit softening enzymes (PE, EGase, exo-PG and endo-PG) in the skin as well as in pulp tissues. The reduction was more pronounced with increased concentrations of 1-MCP. 1-MCP treated fruit showed different rates of fruit softening and activities of ethylene biosynthesis enzymes in the skin and pulp tissues which warrant further investigation on regulation of gene expression related to these enzymes with the inhibitory effect of 1-MCP.     

Volatile profiles of ripening West Indian and Guatemalan-West Indian avocado cultivars as affected by aqueous 1-methylcyclopropene

Separate experiments were conducted with three major commercial avocado (Persea americana Mill.) cultivars grown in Florida: ‘Simmonds’ (early-season, West Indian race); ‘Booth 7’ (mid-season, Guatemalan-West Indian hybrid); and ‘Monroe’ (late-season, Guatemalan-West Indian hybrid). Fruit were harvested at preclimacteric stage and left untreated (Control) or treated 24 h after harvest with aqueous 1-methylcyclopropene (1-MCP) at 1.39 (treatment M1) or 2.77 μmol L⁻¹ a.i. (treatment M2) (75 or 150 μg L⁻¹) for 1 min at 20 °C. Whole fruit ripening was monitored at 20 °C/92% ± 3% R.H. and based on whole fruit firmness, respiration and ethylene evolution. Fruit volatiles were assessed at preclimacteric (24 h after harvest), mid-ripe (half of initial fruit firmness) and ripe maturity stages, from 100 g of chopped pulp using a purge and trap system. Untreated, firmer fruit ‘Monroe’ (268 N at harvest) ripened within 12 d of harvest while softer fruit ‘Simmonds’ (118 N) ripened within only 6 d. 1-MCP treatment extended ripening time from 33% (M1) to 83% (M2). All fruit softened normally, indicating the potential benefits of aqueous 1-MCP as a postharvest treatment for avocado when applied at these concentrations. Volatile profiles differed among the three cultivars with several compounds detected in only one cultivar, results that may contribute to a potential identification of the origin of the cultivar based on fruit volatile composition. The West Indian cultivar ‘Simmonds’ had much higher emission of hexanal (preclimacteric fruit) and cis-3-hexenal and cis-3-hexen-1-ol (ripe fruit) than the Guatemalan-West Indian hybrids ‘Booth 7’ and ‘Monroe’. On the other hand, these latter hybrids had much higher levels of alkanes than ‘Simmonds’. Treatment with 1-MCP increased emissions of alkanes during ripening of ‘Booth 7’ and ‘Monroe’. Total volatiles of avocado decreased during ripening mainly due to the significant reduction of sesquiterpenes, the main group of volatiles in all cultivars at harvest (‘Simmonds’, 53%; ‘Booth 7’, 78%; ‘Monroe’, 66%). β-Caryophyllene was the major compound at harvest, but decreased to less than 2% in ripe fruit, at which point most sesquiterpenes were not detected. Among the 10 sesquiterpenes commonly found in the avocado cultivars in this study, only α-Copaene had significantly higher emissions in mid-ripe fruit treated with the higher concentration of 1-MCP (2.77 μmol L⁻¹ a.i.), suggesting that ethylene participates in the regulation of this sesquiterpene.     

A new strain of Metschnikowia fructicola for postharvest control of Penicillium expansum and patulin accumulation on four cultivars of apple

The efficacy of three antagonistic yeasts, Metschnikowia pulcherrima strain MACH1, M. pulcherrima strain GS9, and Metschnikowia fructicola strain AL27, against Penicillium expansum and patulin accumulation was evaluated on apples stored at room (22 ± 1 °C for 7 days) and cold temperatures (1 ± 1 °C for 56 days). To increase the potential range of application of the biocontrol agents (BCAs), their efficacy was evaluated on four cultivars of apple, i.e. ‘Golden Delicious’, ‘Granny Smith’, ‘Red Chief’ and ‘Royal Gala’. AL27 was more effective than MACH1 and GS9 in the control of blue mold rot and in the reduction of patulin accumulation. The efficacy of AL27 was in most cases similar to the chemical control used, making the antagonist as competitive as chemical fungicides. In vitro experiments showed that AL27 reduced the conidial germination and germ tube length of P. expansum more than the other strains. The three BCAs were more effective in the control of blue mold rot on ‘Golden Delicious’ apples than on the other tested cultivars.     

Measuring consumer response to ‘Gala’ apples treated with 1-methylcyclopropene (1-MCP)

Post-harvest apple treatment with 1-methylcyclopropene (1-MCP) was previously found to inhibit fruit ripening but also to inhibit the production of volatile compounds that contribute to apple flavor. The first objective of this study was to determine if consumers could distinguish 1-MCP treated and untreated Gala apples [Malus sylvestris L. (Mill.) var. domestica Borkh. Mansf.] following long-term storage. Chemical analysis showed 1-MCP treated fruit had reduced flavor volatiles compared to untreated fruit. Consumer difference tests showed they could distinguish between 1-MCP treated and untreated fruit. A second objective was to compare consumers’ acceptance for 1-MCP treated to untreated apples. Both 1-MCP treated and untreated apples received high overall liking scores that were not significantly different. Equal numbers of consumers indicated preference for 1-MCP treated and untreated fruit and there was no difference in purchase intent. However, subsets of consumers who eat Gala, Fuji or Red Delicious apples showed preference for untreated over 1-MCP treated fruit compared to consumers who do not eat these varieties.     

Effects of repeated 1-methylcyclopropene (1-MCP) treatments on ripening and superficial scald of ‘Cortland’ and ‘Delicious’ apples

Postharvest 1-MCP can maintain fruit quality and inhibit development of superficial scald, a physiological storage disorder found in apple fruit, but the extent of the inhibition can vary by cultivar. In this study, we investigated whether multiple applications of 1-MCP, which are now permitted by a label modification of the commercial 1-MCP product, SmartFresh™, might improve scald control. ‘Cortland’ and ‘Delicious’ apples were untreated, treated on the day of harvest with the antioxidant inhibitor of scald, diphenylamine (DPA), or with 1 μL L⁻¹ 1-MCP at different intervals after harvest. Treatment times (days) were 1, 4, 7, 1 + 4, 4 + 7, 1 + 4 + 7, 7 + 14, 7 + 28, 7 + 42, and 7 + 84. Internal ethylene concentrations (IECs), flesh firmness, and accumulations of α-farnesene and conjugated trienols (CTols) were measured at harvest, at the time of treatment, and at intervals during air storage at 0.5 °C for up to 36 weeks. Scald was completely inhibited by DPA and all 1-MCP treatments in ‘Delicious’. However, effective control of scald in ‘Cortland’ was obtained with 1-MCP treatments within the first 4 days of harvest, either alone or in combination. Scald control with delayed 1-MCP treatments resulted in poorer scald control that was comparable to that obtained with DPA. IECs and α-farnesene accumulation were similar in untreated and DPA treated fruit, but inhibited by 1-MCP. However, differences among 1-MCP treatments became more evident with increasing storage periods. Inhibition of IECs and α-farnesene accumulation was greater in fruit treated on days 1, 4, 1 + 4, 4 + 7, 1 + 4 + 7, than on day 7 alone. A second application of 1-MCP on day 14 to fruit treated on day 7 increased inhibition of IECs, α-farnesene and CTol accumulations, but increasing delays before the second 1-MCP treatment resulted in progressively less inhibition of these factors. Similar effects of treatment on IECs, α-farnesene and CTol accumulations were found for both cultivars, even though no scald was detected in treated ‘Delicious’ apples. The results indicate that initial 1-MCP treatments should be applied to faster ripening cultivars such as ‘Cortland’ within a few days of harvest.     

Ethylene biosynthesis in apricot: Identification of a ripening-related 1-aminocyclopropane-1-carboxylic acid synthase (ACS) gene

Apricots are climacteric fruits with a high susceptibility to flesh softening and loss of flavor during postharvest storage, and most of the ripening processes are regulated by ethylene, which also has an effect on its own biosynthesis. To understand this process in apricot, inhibition of ethylene biosynthesis and perception was performed for studying key genes involved in the ethylene biosynthetic pathway. Apricots, cv. “Patterson”, were harvested with yellow-green ground color and immediately treated with either the ethylene perception inhibitor 1-methyl cyclopropene (1-MCP) at 10 μL L⁻¹ or the ethylene biosynthesis inhibitor aminoethoxyvinylglycine (AVG) at 1 g L⁻¹. After treatment, quality and physiological attributes such as firmness, color, total soluble solids, acidity, fruit weight, ethylene production and respiration rates were evaluated every 2 d until they ripened at 20 °C. Gene expression analysis was performed by quantitative polymerase chain reaction (qPCR). Both ethylene inhibitors were effective in reducing ethylene production, respiration rate and fruit softening. Three 1-aminocyclopropane-1-carboxylic-acid synthase (ACS) genes were characterized, but only the expression of ACS2 was highly reduced by ethylene inhibition, suggesting a key role in ethylene synthesis at ripening. Contrarily, ACS1 and ACS3 showed a higher expression under ethylene inhibition suggesting that the corresponding genes are individually regulated in a specific mode as observed in other climacteric fruits. Finally, changes in 1-aminocyclopropane-1-carboxylic-acid oxidase genes did not show a consistent pattern of ethylene modulation.     

Effect of 1-methylcyclopropene on ripening of ‘Canino’ apricots and ‘Royal Zee’ plums

‘Canino’ apricots and ‘Royal Zee’ plums were treated with 1000 nl l⁻¹ 1-methylcyclopropene (1-MCP) at 20 °C for 20 h following harvest before 0 °C storage. After 5 days storage for apricots and 10 days for plums and after 30 days storage for both, fruit were moved to 20 °C for ripening. In addition, apricots were stored for 20 days and then treated with 1-MCP concentrations of 10, 100 and 1000 nl l⁻¹ at removal and held for ripening. Ethylene production and respiration rate, as well as fruit quality of apricots varied with treatment. Ethylene production was efficiently inhibited by 1000 nl l⁻¹ 1-MCP in fruit treated after storage but not in fruit treated before storage. Fruit softening was associated with ethylene production and affected by 1-MCP in a concentration dependent manner when treated after storage, while 1-MCP did not affect softening in prestorage treated fruit. The color change of fruit was ethylene-independent and not affected by 1-MCP. Internal flesh browning was decreased by 1-MCP regardless of the concentration when treated after storage, while it was enhanced in fruit treated before storage. Decay development in apricots was decreased by 1-MCP in a concentration dependent manner. Ethylene production and respiration in ‘Royal Zee’ plums was greatly inhibited by 1-MCP during ripening after both short-term (10 day) and long term (30 day) storage. Parameters associated with ripening processes were decreased significantly by 1-MCP, including softening, color change, and loss of titratable acidity. These data demonstrate that 1-MCP has potential to delay ripening of apricots and plums, but the cultivar, maturity of fruit, and time of application must be chosen carefully. It is suggested that 1-MCP is more efficient for extending the shelf life and improving the quality of ‘Canino’ apricots directly marketed or after storage, whereas it might be a potent compound for extending both storage period and shelf life of ‘Royal Zee’ plums.     

‘苏帅’苹果表观遗传变异分析

[目的]为探索杂交品种‘苏帅’表观优良性状遗传的生理机制。[方法]以杂交品种‘苏帅’和父本‘金冠’、母本‘印度’为材料, 通过分析 ‘苏帅’和‘金冠’、‘印度’等苹果品种的节间数、节间长度以及叶片内源激素、果实可溶性固形物、可滴定酸、叶绿素的含量; 采用石蜡切片法、扫描电镜等方法比较其叶片结构, 测定了叶片光合气体交换参数。[结果]结果表明, ‘金冠’和‘印度’的节间长度显著大于‘苏帅’苹果的节间长度,‘苏帅’苹果叶片叶绿素相对含量显著高于‘金冠’和‘印度’; 果实可溶性固形物含量与‘金冠’虽然没有显著性差异, 但可滴定酸的含量明显高于‘金冠’苹果; ‘苏帅’苹果内源激素脱落酸(ABA)、赤霉素(GA)、吲哚－3－乙酸(IAA)、玉米素核苷(ZR)与‘金冠’和‘印度’相比均呈显著差异。‘苏帅’苹果叶片下表皮气孔长度大于‘印度’, 且差异极显著,但小于‘金冠’,气孔宽度显著宽于‘印度’,与‘金冠’苹果没有明显差异, ‘苏帅’苹果气孔密度极显著小于‘金冠’和‘印度’;‘金冠’和‘印度’栅栏细胞组织排列较紧密, 栅栏组织和海绵组织分界限明显,而‘苏帅’苹果的栅栏组织细胞排列相对杂乱,细胞大小也不一致, 与海绵组织界限不明显; ‘苏帅’苹果叶片、上下表皮、栅栏组织及海绵组织厚度显著高于‘金冠’和‘印度’,栅栏组织和海绵组织比值也显著高于‘金冠’和‘印度’。[结论] ‘苏帅’苹果与父母本相比表现出节间短、叶片变厚、光合作用增强等性状多样性, 为苹果生产提供了新品种,也为苹果新品种的培育提供了珍贵的种质资源。     

Responses of 1-MCP application in plums stored under air and controlled atmospheres

The potential of 1-MCP for controlling ripening in ‘Angeleno’ plum fruit under air and controlled atmosphere (CA) storage was explored, and the possibility that 1-MCP can inhibit development of brown rot caused by Monilinia laxa and internal breakdown in ‘Fortune’ and ‘Angeleno’ plums tested. After harvest, fruit were exposed to 300 and 500 nl l⁻¹ (in 2003) and 500 nl l⁻¹ 1-MCP (in 2004) at low temperatures (0–3 °C) for 24 h. After treatment the plums were stored in air at 0 °C and ‘Angeleno’ fruit were also stored in CA storage (1.8% O₂ + 2.5% CO₂). Following storage, fruit were kept at 20 °C. In ‘Angeleno’ fruit, 1-MCP was effective in delaying the loss of firmness and colour changes during holding at 20 °C. 1-MCP reduced brown rot in fruit stored in CA but no significant reduction was found in air storage. Internal breakdown, a major physiological storage disorder in plums, was inhibited by 1-MCP treatment. Furthermore, since 1-MCP applied in air storage showed better results than the control in CA conditions, an application of 1-MCP before air storage could be the best way to reduce the ripening process for short or medium storage periods (40 and 60 days). CA storage plus 1-MCP treatment could be used for long periods (80 days).     

Ethylene synthesis,ripening capacity,and superficial scald inhibition in 1-MCP treated ‘d’Anjou’ pears are affected by storage temperature

A continuing challenge for commercializing 1-methylcyclopropene (1-MCP) to extend the storage life and control superficial scald of ‘d’Anjou’ pear (Pyrus communis L.) is how to initiate ripening in 1-MCP treated fruit. ‘D’Anjou’ pears harvested at commercial and late maturity were treated with 1-MCP at 0.15 μL L⁻¹ and stored either at the commercial storage temperature −1.1 °C (1-MCP@−1.1 °C), or at 1.1 °C (1-MCP@1.1 °C) or 2.2 °C (1-MCP@2.2 °C) for 8 months. Control fruit stored at −1.1 °C ripened and developed significant scald within 7 d at 20 °C following 3–5 months of storage. While 1-MCP@−1.1 °C fruit did not develop ripening capacity due to extremely low internal ethylene concentration (IEC) and ethylene production rate for 8 months, 1-MCP@1.1 °C fruit produced significant amounts of IEC during storage and developed ripening capacity with relatively low levels of scald within 7 d at 20 °C following 6–8 months of storage. 1-MCP@2.2 °C fruit lost quality quickly during storage. Compared to the control, the expression of ethylene synthesis (PcACS1, PcACO1) and signal (PcETR1, PcETR2) genes was stable at extremely low levels in 1-MCP@−1.1 °C fruit. In contrast, they increased expression after 4 or 5 months of storage in 1-MCP@1.1 °C fruit. Other genes (PcCTR1, PcACS2, PcACS4 and PcACS5) remained at very low expression regardless of fruit capacity to ripen. A storage temperature of 1.1 °C can facilitate initiation of ripening capacity in 1-MCP treated ‘d’Anjou’ pears with relatively low scald incidence following 6–8 months storage through recovering the expression of certain ethylene synthesis and signal genes.