Identification of organic acid-related genes and their expression profiles in two pear (Pyrus pyrifolia) cultivars with difference in predominant acid type at fruit ripening stage

‘Yandangxueli’ is a pear cultivar with predominant citric acid in the ripe fruit, different from most of pear cultivars such as ‘Gengtouqing’ in which malic acid is the predominant acid type. It was found that ‘Yandangxueli’ accumulated citric acid for three times against that in ‘Gengtouqing’ at fruit ripening stage. To investigate the mechanism of citric acid accumulation in ‘Yandangxueli’, organic acids content, gene expression and enzyme activity were studied in both cultivars. Five genes, Pp:mtCs, Pp:cyAco, Pp:cyIdh, Pp:mtMdh and Pp:cyMe which encoded citric synthase (CS), cytosolic aconitase (cyACO), NADP-dependent isocitrate dehydrogenase (NADP-IDH), NAD-dependent malate dehydrogenase (NAD-MDH) and NADP-dependent malic enzyme (NADP-ME) respectively, were identified from pear fruit. Their expression profiles and the corresponding enzyme activities were determined throughout fruit development in both cultivars. Results from these enzymes indicated that there were no strict relationship between gene expression, enzyme activity and citric acid accumulation. Expression analysis for two Py:vVAtp genes encoding vacuolar H⁺-ATPase A subunit and one Py:vVpp gene encoding Vacuolar H⁺-pyrophosphatase showed that they were all with up-regulated expression at the later development stage of ‘Yandangxueli’ but with down-regulated expression in ‘Gengtouqing’. Therefore, it is concluded that the different ability in citric acid transportation and storage might be involved in the high citric acid content in ‘Yandangxueli’.     

Effect of distance from early flowering pollinizers ‘Maypole’ and ‘Dolgo’ on ‘Fiji’ fruit set

An apple orchard consisting of a single cultivar under the condition of natural pollination must have suitable pollinizers such as Crab apples to ensure stable fruit production. We selected ‘Maypole’ and ‘Dolgo’ as pollinizers for the cultivar ‘Fuji’, and investigated the rate of fruit and seeds in ‘Fuji’ fruits produced by pollen of the pollinizers. We developed a method for tracing pollen flow based on the leaf color of progeny and S-RNase allele of ‘Maypole’, and on Simple Sequence Repeat (SSR) analyses of ‘Maypole’ and ‘Dolgo’. These were powerful tools for determining the distance insects (mainly Osmia cornifrons) carry pollen from the pollinizers to ‘Fuji’. Although the fruit set of ‘Fuji’ apples was not reduced with increasing distance between ‘Fuji’ apple trees and pollinizers (probably due to pollen flow from other commercial cultivars planted outside the area), the rate of ‘Fuji’ apple fruit produced by the pollen of the pollinizers decreased with increasing distance. The rate of fruit produced by the pollinizers was 84% and 77% when ‘Fuji’ was 2.5 m from ‘Maypole’ and ‘Dolgo’, respectively, and 71% and 64% when ‘Fuji’ was 5 m from ‘Maypole’ and ‘Dolgo’, respectively, but was reduced to 47% and 39% when ‘Fuji’ was 10 m from ‘Maypole’ and ‘Dolgo’, respectively. However, the spacing could cause reduced fruit size, and require extra fruit thinning for producing large fruits. It could also cause reduction of the yield for increasing the planting area of pollinizers. We recommend that pollinizers should be planted not more than 10 m from ‘Fuji’.     

Induction of disease resistance and ROS metabolism in navel oranges by chitosan

The objective of this work was to evaluate how disease resistance and reactive oxygen species (ROS) metabolism in harvested navel oranges (Citrus sinensis L. Osbeck) may be affected by chitosan. Fresh navel oranges were treated with 2% chitosan or 0.5% glacial acetic acid (control) solution for 1 min, and some were inoculated with Penicillium italicum and Penicillium digitatum. Then, the fruit were stored at 20 °C and 85–95% RH. Treatment with 2% chitosan significantly reduced the disease incidence and the lesion diameter compared with control fruit. This treatment effectively enhanced the activities of peroxidase (POD) and superoxide dismutase (SOD), and levels of glutathione (GSH) and hydrogen peroxide (H₂O₂), inhibited the activities of catalase (CAT) and the decreases of ascorbate (AsA) content during navel orange fruits storage. Ascorbate peroxidase (APX) activity in the navel orange fruit was induced slightly by the chitosan treatment during 14–21 days storage. However, glutathione reductase (GR) activity in the fruit was not enhanced by the chitosan treatment. These results indicated that chitosan treatment could induce the navel orange fruit disease resistance by regulating the H₂O₂ levels, antioxidant enzyme and ascorbate–glutathione cycle.     

QTL analysis and regression model for estimating fruit setting in young Citrus trees based on molecular markers

This study was conducted to assess the genetic basis and develop a regression model for a QTL trait, fruit setting of a full sib population of 164 hybrids obtained between ‘Clementine’ mandarin (Citrus reticulata Blanco ‘Clementine) and ‘Orlando’ tangelo’ (Citrus paradisi Macf. ‘Duncan’ × C. reticulata Blanco ‘Dancy’). Fruit setting of a 164 full sib population field-planted in 2007 was evaluated by visual counting of fruits in 2008, 2009, and 2010. We estimated linkage groups and effects of QTLs by using MapQTL5. Based on interval mapping, seven linkage groups of the maternal and eight linkage groups of the paternal maps harboured QTLs of the fruits setting, covering a total 300 of 1744 cM Citrus map. Only three segments were associated with all three-year data (one in ‘Clementine’ and two in ‘Orlando’ map) spanning 60 cM of 1744 cM of Citrus linkage map. Twenty-seven (17%) and 13 (8%) hybrids consistently produced less than 5 fruits and the highest number of fruits (≥50) in their 2nd, 3rd, and 4th ages. Based on BACKWARD elimination procedure of PROC REG option nested in SAS program, regression models constructed for 2008, 2009, and 2010s fruit setting explained 35, 81, and 38% of the total phenotypic variation (R²) observed in the 164 full-sib hybrids, respectively (P ≤ 0.05). This study indicated that early fruit setting was a complex trait affected by many QTLs and the regression model developed in this study might be used to predict performances of hybrids with similar genetic background.     

Tree performance and fruit yield and quality of ‘Okitsu’ Satsuma mandarin grafted on 12 rootstocks

The citriculture in Brazil, as well as in other important regions in the world, is based on very few mandarin cultivars. This fact leads to a short harvest period and higher prices for off-season fruit. The ‘Okitsu’ Satsuma (Citrus unshiu Marc.) is among the earliest ripening mandarin cultivars, and it is considered to be tolerant to citrus canker (Xanthomonas citri subsp. citri Schaad et al.) and to citrus variegated chlorosis (Xylella fastidiosa Wells et al.). Despite having regular fruit quality under hot climate conditions, the early fruit maturation and absence of seeds of ‘Okitsu’ fruits are well suited for the local market in the summer (December through March), when the availability of citrus fruits for fresh consumption is limited. Yet, only a few studies have been conducted in Brazil on rootstocks for ‘Okitsu’. Consequently, a field trial was carried out in Bebedouro, São Paulo State, to evaluate the horticultural performance of ‘Okitsu’ Satsuma mandarin budded onto 12 rootstocks: the citrandarin ‘Changsha’ mandarin (Citrus reticulata Blanco) × Poncirus trifoliata ‘English Small’; the hybrid Rangpur lime (Citrus limonia Osbeck) × ‘Swingle’ citrumelo (P. trifoliata (L.) Raf. × Citrus paradisi Macfad.); the trifoliates (P. trifoliata (L.) Raf.) ‘Rubidoux’, ‘FCAV’ and ‘Flying Dragon’ (P. trifoliata var. monstrosa); the mandarins ‘Sun Chu Sha Kat’ (C. reticulata Blanco) and ‘Sunki’ (Citrus sunki (Hayata) Hort. ex. Tanaka); the Rangpur limes (C. limonia Osbeck) ‘Cravo Limeira’ and ‘Cravo FCAV’; ‘Carrizo’ citrange (Citrus sinensis × P. trifoliata), ‘Swingle’ citrumelo (P. trifoliata × C. paradisi), and ‘Orlando’ tangelo (C. paradisi × Citrus tangerina cv. ‘Dancy’). The experimental grove was planted in 2001, using a 6 m × 3 m spacing, in a randomized block design. No supplementary irrigation was applied. Fruit yield, canopy volume, and fruit quality were assessed for each rootstock. A cluster multivariate analysis identified three different rootstock pairs with similar effects on plant growth, yield and fruit quality of ‘Okitsu’ mandarin. The ‘Flying Dragon’ trifoliate had a unique effect over the ‘Okitsu’ trees performance, inducing lower canopy volume and higher yield efficiency and fruit quality, and might be suitable for high-density plantings. The ‘Cravo Limeira’ and ‘Cravo FCAV’ Rangpur limes induced early-ripening of fruits, with low fruit quality. ‘Sun Chu Sha Kat’ and ‘Sunki’ mandarins and the ‘Orlando’ tangelo conferred lower yield efficiency and less content of soluble solids for the latter rootstock.     

Yield performance,quality characteristics and fruit storability of winter squash cultivars in sub-humid areas

This research investigated the quality traits of eight winter squash cultivars (Cucurbita maxima, C. moschata, C. pepo and interspecific hybrids of C. moschata × C. pepo) during three years, but only ‘Tetsukabuto’ and ‘Violina’ were tested each year. In 2005 these two varieties were compared to ‘Butternut’; in 2006 to ‘Red Kury’, ‘Tan Cheese’ and ‘Kabosha’; in 2007 to ‘Red Kury’, ‘Mooregold’, and ‘Winter Luxury’. The characteristics recorded were: fruit yield, storage ability, chemical composition and sensory quality. Following harvest and sorting of marketable winter squash, 50 fruits for each cultivar were stored at 12 °C for 12 weeks the 1st year, and for over 20 weeks in 2nd and 3rd years, during which rotten fruits were counted. Compositional analyses regarding sugar concentration, starch, carotenoids and dry matter content at harvest and after storage were carried out; a panel test was organised to assess sensory traits. The most interesting cultivars were ‘Tetsukabuto’ for yield, ‘Tetsukabuto’ and ‘Mooregold’ for storability, ‘Red Kury’ and the same ‘Tetsukabuto’ particularly for soluble sugars and carotenoids and sensory appreciation.     

Changes in starch and soluble sugar concentrations in winter squash mesocarp during storage at different temperatures

The effects of storage at 5, 10 or 15 °C for 6 months on the concentrations of starch and soluble sugar in winter squash (Cucurbita maxima Duch.) cultivar ‘TC2A’ fruits were examined. Starch contents were significantly lower at 15 °C than at the other temperatures, although concentrations decreased throughout the storage period at all temperatures. Total soluble sugar contents increased during the first 3 months of storage regardless of temperature, and decreased at 5 °C or 15 °C, but not at 10 °C after 6 months. Myo-inositol and raffinose concentration patterns were more complex, and may reflect some role in regulating fruit metabolism during storage that may be important in maintaining overall squash fruit quality.     

Changes in organic acids and acid metabolism enzymes in melon fruit during development

A novel melon cultivar (‘Flavor No. 3’) containing high acid concentration as well as high sugar content in the mature fruit was successfully bred out. The aim of this study is to determine the main organic acids present in this novel melon and to clarify the mechanisms of acid accumulation. Fruit development and acid accumulation patterns in ‘Flavor No. 3’ were compared with those of a conventional low-acid melon, ‘Xuelihong’. The titratable acidity (TA) and organic acids in fruits were measured at different developmental stages. There is a positive correlation between TA and citric acid content in ‘Flavor No. 3’, indicating that citric acid is the predominant organic acid present throughout the fruit development. The activities of the enzymes involved in citric acid metabolism, including phosphoenolpyruvate carboxylase (PEPC), citrate synthase (CS), aconitase (ACO), isocitrate dehydrogenase (IDH), malate dehydrogenase (MDH), and malic enzyme (ME), were determined. Although no single enzyme was responsible for citric acid accumulation throughout the fruit development, the activities of ACO and ME were positively correlated with citric acid concentration in the two cultivars from 5 to 25 days after pollination (DAP). Moreover, ACO and ME activities in ‘Flavor No. 3’ increased over time and were higher than those in ‘Xuelihong’, resulting in citric acid accumulation in the fruits of ‘Flavor No. 3’. Although a sharp decrease in ACO activity took place after 25 DAP in the fruits of ‘Flavor No. 3’, the earlier accumulation of citric acid was sufficient enough to maintain the high acid concentration until the melon fruits became mature.     

Differences in fruit colour development,anthocyanin content,fruit quality and consumer acceptability of eight ‘Gala’ apple strains

The effect of strain on fruit colour development, chromaticity values and anthocyanin content, fruit quality and consumer acceptance was evaluated on eight ‘Gala’ apple strains (Malus domestica Borkh.) at the IRTA (Spain), during a period from 4 weeks before to 1 week after commercial harvest. Based on fruit colour, measured with a Minolta CR-200 portable tristimulus colorimeter, and anthocyanin content, the most coloured strains were ‘Royal Beaut’, ‘Buckeye Gala’ (both semistriped) and ‘Ruby Gala’ (blushed). ‘Brookfield’ and ‘Schniga’ (both striped) provided an intermediate level of colour, while ‘Galaxy’ and ‘Mondial Gala’ (both striped) were the least coloured ones. The most important increase in colour development was recorded from 2 weeks before the commercial harvest and continued also increasing after harvest. Highly coloured strains developed a red colour on both fruit sides with greater average of fruit surface coloured, while the less coloured strains exhibited different colouration between sides, more bicolour fruits and lower average of fruit coloured.     

Changes of physicochemical quality,phenolics and vitamin C content in hardy kiwifruit (Actinidia arguta and its hybrid) during storage

The effect of storage and the stage of maturity of hardy kiwifruits on the physicochemical quality, phenolics (TPC) and ascorbic acid content (AAC), as well as antioxidant activity (AA) were studied in this work. The phenolic compounds in hardy kiwifruits were separated and characterized by HPLC. The investigation was carried out on the two cultivars of Actinidia arguta – ‘Weiki’, ‘74-49’ and the hybrid of A. arguta and Actinidia purpurea (‘D14’). Fruit firmness rapidly decreased and the soluble solid content (SSC) increased for all cultivars during the first 14 days of storage at 1 °C. The AAC and TPC in vine ripe fruits were similar to the ones of the fruits of storage harvest maturity (8–10% SSC). AA content depended on the clone and either decreased during storage or remained almost unchanged. There was an increase in TPC after 7 days of keeping the fruits in a cold store chamber at the temperature 1 °C, but a longer period of storage caused a decrease in these compounds. AA (at harvest for storage purposes) was higher than that of vine ripe fruits and the ability to absorb free radicals slightly decreased during storage. There was a strong correlation between AAC, TPC and AA. That means that phenolics and vitamin C affect the antioxidant activity of hardy kiwifruits.     

Biochemical changes associated with hardening of endocarp in Spondias mangifera Willd. fruits during developmental stages

Spondias mangifera is a unique, fleshy, drupe fruit with a mango flavor. The hardening of endocarp during development and maturation of hog plum is a major bottleneck, which reduces the edibility of the fruits by 80% and makes them unmarketable. For the first time, three distinct maturity stages were demonstrated. Distinct pattern in change in surface color, accumulation of carotene pigments and biochemical components in fruits were observed. The decrease in chlorophyll ‘a’ and chlorophyll ‘b’ correlated well with the downhill trend in Transmittance Refractory Spectroscopy (TRS) – ‘L’, ‘a’, ‘b’ values. The decrease in the total chlorophyll along with the accumulation of carotene pigment contributes significantly to the lightness of surface color and reflected in ‘L’ hue values. Confluence of carotene and total chlorophyll on the 9th week provides a distinct marker for maturity. Complete convergence of five carpells on the 9th week after fruit set, heralds the onset of maturation in Indian hog plum and which was adjudged as visual index of maturity. Hardening of endocarp begins after attainment of optimum maturity and last for 2 weeks. Pre-climacteric plunge in respiration along with coordinated changes in pigments, and accumulation of ascorbic acid, phenols, presets the optimum maturity period of 9th week after fruit set. Pre-climacteric falls in respiration with lowest output of 40.57 mg CO₂/kg-h concentration on 9th week after fruit set indicates the attainment of fruit maturity. Harvesting of hog plum between 9th and 11th weeks provides the fruit with soft endocarp and complete edibility.     

Effect of synthetic auxins on fruit size of five cultivars of Japanese plum (Prunus salicina Lindl.)

Most of the Japanese plum (Prunus salicina) cultivars grown in Israel produce relatively small fruit. Application of 2 l solution tree⁻¹ of 25 mg l⁻¹ 2,4-dichlorophenoxypropionic acid (2,4-DP) as butoxyethyl ester (Power™), 15 mg l⁻¹ 3,5,6-trichloro-2-pyridyloxyacetic acid (3,5,6-TPA) as free acid (Maxim^®), or 25 mg l⁻¹ 2,4-dichlorophenoxyacetic acid (2,4-D) + 30 mg l⁻¹ naphthaleneacetic acid (NAA) (0.3% Amigo™) at the beginning of pit-hardening, when fruitlet diameter was ca. 22 mm, caused an appreciable and significant increase in fruit size. The yield of large fruit per cv.: ‘Kesselmen’ (100% increase), ‘Songold’ (100%), ‘Black Diamond’ (800%), ‘Royal Diamond’ (160%) and ‘Royal Zee’ (100%). As a result, the total yield of all five cultivars was also increased dramatically. Anatomical studies with ‘Songold’ revealed that the main effect of these synthetic auxins was via direct stimulation of fruit cell enlargement. The above auxins had no negative effect either on fruit quality at harvest (and after 1 week in shelf-life), or on return yield in the following year.     

Grafting effects on postharvest ripening and quality of 1-methylcyclopropene-treated muskmelon fruit

In addition to managing soil-borne diseases in muskmelon (Cucumis melo L.) production, grafting with resistant rootstocks may impact fruit quality. The ethylene antagonist 1-methylcyclopropene (1-MCP) has been shown to extend shelf life of fresh muskmelon fruit. Postharvest characteristics of 1-MCP-treated melon fruit as affected by grafting, however, have not been well examined. This study was conducted to explore the influence of grafting with different rootstocks on ripening and quality attributes of 1-MCP-treated muskmelon fruit during postharvest storage. Grafted ‘Athena’ muskmelon with two commercial squash interspecific hybrid rootstocks including ‘Strong Tosa’ and ‘Tetsukabuto’ as well as non-grafted and self-grafted ‘Athena’ were grown in replicated field plots at the University of Florida Plant Science Research and Education Unit (Citra, FL, USA) during April–June 2010. Half-slip fruit from two harvests were treated with 1.0 μL L⁻¹ 1-MCP (18 h, 20 °C) and analyzed during storage at 13 °C. For fruit from the 27 May harvest, whole fruit and mesocarp firmness, titratable acidity, soluble solids, and ascorbic acid content were measured, while production of ethylene and CO₂ was determined on fruit from the 29 June harvest. Grafting did not show a significant impact on fruit yield but affected the fruit shelf life significantly. Fruit from non-grafted ‘Athena’ and ‘Athena’ grafted onto ‘Strong Tosa’ demonstrated a shelf life of 31 d for the first harvest and 22 d for the second harvest. Shelf life of fruit from self-grafted ‘Athena’ and ‘Athena’ grafted onto ‘Tetsukabuto’ declined by 6 d and 3 d for the first and second harvest, respectively. Whole fruit firmness decreased by approximately 15.5% on average from 13 to 31 d except day 19 as a result of grafting, but to a lesser extent with ‘Strong Tosa’ rootstock. Mesocarp firmness of grafted melon was reduced by about 30.2% at days 13 and 19 compared to non-grafted ‘Athena’ fruit. In contrast, titratable acidity, soluble solid content, and ascorbic acid concentration were less affected by grafting. All the measurements except for ethylene and CO₂ production declined during storage regardless of the grafting treatment. Compared with ‘Strong Tosa’ rootstock, ‘Tetsukabuto’ resulted in a more rapid ripening under 1-MCP application, as reflected by earlier increase in ethylene production and higher respiratory rate. The study demonstrates that grafting effects on postharvest ripening and quality of ‘Athena’ muskmelon can vary markedly with rootstocks used.     

Acid invertase as a serious candidate to control the balance sucrose versus (glucose + fructose) of banana fruit during ripening

Six banana varieties: 3 ‘dessert’ ones: ‘IDN 110’; ‘Kirun’; and ‘Grande Naine’, and 3 ‘cooking’ ones: ‘Galéo’; ‘Sowmuk’; and ‘French’ were used to investigate the relationship between sugar profiles and activities of sucrose-phosphate synthase (SPS, EC 2.4.1.14), sucrose synthase (SuSy, EC 2.4.1.13) working in the hydrolytic way, invertases (EC 3.2.1.26) (neutral (NIV) and acid (AIV)). Expression of a Musa cell-wall invertase (MaCwINV1/pBANUU103) gene was additionally studied during fruit development and ripening ex planta after ethylene treatment of two of these varieties. Close amounts of soluble sugars (sucrose + glucose + fructose) were measured in the different varieties at ripe stage. SPS activity was either almost constant or increased continuously or transitorily during ripening of all varieties, concomitantly to total soluble sugar (sucrose + glucose + fructose) accumulation. Together with previous data obtained from ‘Cavendish’, our data lead us, (i) to strengthen the hypothesis that this enzyme is likely to have a major role in the synthesis of sucrose during ripening of banana and (ii) to underline the complexity of the mode of SPS activity regulation already pointed out. Interestingly, for the first time in banana, two diploid and cooking varieties: ‘Galéo’ and ‘Sowmuk’ were found almost sucrose-free, in good agreement with a 6.4-fold higher mostly vacuolar AIV activity than that of the two desserts and diploid ones. Otherwise, expression of a MaCwINV1/pBANUU103 (cell wall) gene was followed in the two most contrasted varieties in matter of sucrose content: ‘Sowmuk’ almost sucrose-free, and ‘IDN 110’ accumulating high level of sucrose. Compared to ‘IDN110’, the mRNA level of MaCwINV1/pBANUU103 gene was higher (up to 173-fold) in ‘Sowmuk’ concomitantly with the low level of sucrose of ‘Sowmuk’. Our data let us to conclude that the AIV is probably one of the main determinants involved in the regulation of sucrose level during banana fruit ripening, even if the form, vacuolar or cell wall-linked is not determined yet. Otherwise, the MaCwINV1/pBANUU103 gene appears as a putative candidate gene that could contribute to regulate this level.     

Yield and fruit quality of ‘Gala’ and ‘Fuji’ apple trees protected by white anti-hail net

Covering apple orchards with nets protects them from hailstorms but this changes the amount and quality of the light supplied to the trees. This study was carried out to assess the effects of shade provided by white protection net on yield and fruit quality of apple orchards in Southern Brazil, along three growing seasons (2002/2003, 2003/2004, and 2004/2005). ‘Gala’ and ‘Fuji’ apple trees (on MM111 rootstock, raised as slender spindles, with a spacing of 5.5 m × 3 m) were left uncovered (control trees) or covered with white net. The white net reduced the photosynthetically active radiation (λ = 400–700 nm) accumulated over the trees’ canopy along a day by 18.4%, leading to a higher leaf chlorophyll content in both cultivars, and a higher specific leaf area in ‘Gala’. The shade provided by the net did not affect yield and increased fruit average weight in both cultivars. At harvest, the number of seeds per fruit and severity of russet were not affected by netting in both cultivars, as well as the incidence of watercore in ‘Fuji’. The main beneficial effects of the white net were the reduced incidence of sunburn on ‘Gala’, reduced incidence of bitter pit during cold storage in both cultivars, and reduced incidence of apple scab at harvest and of decay after cold storage in ‘Fuji’. Fruit fly damage in ‘Gala’ (assessed in one season) was reduced by netting. However, netting affected negatively some fruit quality attributes. ‘Gala’ and ‘Fuji’ apples from trees protected by the net had a poor skin color (pale blush and a more intense green background color) at harvest. The net reduced flesh firmness and SSC, and increased the starch index of ‘Gala’ apples at harvest, and reduced flesh firmness and SSC, assessed after cold storage and shelf life, in fruits of both cultivars.     

Relationship between early watercore development and leaf photosynthesis or partitioning of photosynthates in apple

Seasonal changes in leaf photosynthetic rate and partitioning of ¹³C taken up by leaves were compared in relation to the development of early watercore using susceptible ‘Orin’ and resistant ‘Fuji’ apples (Malus domestica Borkh.). There was no difference between the cultivars in leaf photosynthetic rate from mid-July to mid-October. The extent of watercore in ‘Orin’ increased from mid-July and peaked in late-August, followed by a decrease through mid-October, while no watercore was detected during the experimental period in ‘Fuji’. Fruit fresh weight and soluble solids content (SSC) were higher in ‘Orin’ than ‘Fuji’ during the earlier stages. Leaf dry weight of ‘Orin’ was higher than that of ‘Fuji’ throughout the experimental period. Accumulation of dry matter in the fruit increased in both cultivars and ‘Orin’ kept significantly higher level than ‘Fuji’ from mid-July to mid-October. The amount of ¹³C distributed to ‘Orin’ fruit at 5 days after labeling was significantly higher than ‘Fuji’ in mid-July just before watercore started to occur. ¹³C taken up by ‘Orin’ fruit with prevalent watercore was also higher than ‘Fuji’ in late-August but the difference was not significant. In mid-October when early watercore was gradually reducing, ¹³C recovered by ‘Orin’ fruit decreased and was significantly lower than ‘Fuji’. The difference of partitioning pattern of ¹³C between the cultivars was associated with the susceptibility to early watercore as well as the changes in fruit growth and SSC. The seasonal changes in the amount of ¹³C distributed to ‘Orin’ fruit were also related to the fluctuations in watercore severity. The results suggest that active phloem transport might be the cause of early watercore in our previous work, which might be induced by the increased partitioning of assimilates to the fruit during summer in susceptible ‘Orin’ compared to resistant ‘Fuji’ and not by the higher potential of leaf photosynthesis.     

Evaluation of ‘Hamlin’ sweet orange + ‘Montenegrina’ mandarin somatic hybrid for tolerance to Xanthomonas axonopodis pv. citri and Xylella fastidiosa

Asiatic citrus canker (ACC), caused by Xanthomonas axonopodis Starr & Garces pv. citri (Hasse) Vauterin et al., and citrus variegated chlorosis (CVC), caused by Xylella fastidiosa Wells et al., are considered the main diseases affecting sweet orange scion varieties in Brazil. Among commercial varieties, mandarins and tangerines are recognized as tolerant to these pathogens. We report herein the production of ‘Hamlin’ sweet orange (Citrus sinensis L. Osbeck) + ‘Montenegrina’ mandarin (Citrus deliciosa Ten.) allotetraploid somatic hybrid plants by protoplast fusion with improved disease tolerance that could be used as a donor of resistance genes in interploid hybridisation. Somatic hybridisation was confirmed by leaf morphology, flow cytometry and RAPD analyses. The somatic hybrid was propagated by grafting and cultivated in a screenhouse for tolerance assays. For X. axonopodis pv. citri assays, buds were collected from both ‘Hamlin’ sweet orange and the somatic hybrid and grafted onto ‘Cleopatra’ mandarin (Citrus reshni hort. ex Tanaka). As a negative control, buds from ‘Mexerica Tardia’ mandarin (C. deliciosa) were collected and grafted onto ‘Cleopatra’ mandarin. Two-month old plants with at least one young vegetative flush were individually spray-inoculated with a 10⁶ CFU mL⁻¹X. axonopodis pv. citri suspension and incubated in a growth chamber, at 27 °C, under 16-h photoperiod. The somatic hybrid showed a statistically significant reduction in susceptibility to ACC 30 days after inoculation. Compared to ‘Hamlin’ sweet orange, disease severity was reduced by 70%, with similar tolerance to that of the mandarin negative control. For X. fastidiosa assays, buds were collected from the somatic hybrid and its parental plants and grafted onto ‘Rangpur’ lime (Citrus limonia Osbeck). The developed plants were needle-inoculated with a X. fastidiosa suspension (8.7 × 10¹⁰ CFU mL⁻¹) into the new growth flush stem. Bacterial population was quantified both at 4 (at the inoculation point) and 8 months (50 cm above the inoculation point) after inoculation. The first evaluation detected X. fastidiosa in 63% of ‘Hamlin’ sweet orange and ‘Hamlin’ + ‘Montenegrina’ mandarin samples. In the second evaluation, X. fastidiosa was detected in 47.4% of ‘Hamlin’ sweet orange and 10.5% of ‘Hamlin’ + ‘Montenegrina’ somatic hybrid samples, suggesting that bacterial movement was restricted in the somatic hybrid. X. fastidiosa was not detected in both evaluations in samples collected from leaves of ‘Montenegrina’ mandarin. These results indicate that the ‘Hamlin’ sweet orange + ‘Montenegrina’ mandarin somatic hybrid has potential for improved disease tolerance that should enhance its value regarding future use in citrus breeding programs.     

H2O2 metabolism during sweet orange (Citrus sinensis L. Osb.) ‘Hamlin’ Xanthomonas axonopodis pv. citri interaction

Sweet orange (Citrus sinensis L. Osb.) ‘Hamlin’ is a canker (Xanthomonas axonopodis pv. citri: Xac) susceptible citrus genotype grown commercially worldwide. Canker causes severe economic losses and restricts the marketability of crop for export. Little is known about the role of oxidative stress in canker development. In the present investigation, sweet orange ‘Hamlin’ leaves were artificially inoculated with Xac to determine the impact of Xac infection on hydrogen peroxide (H₂O₂) metabolism. Characteristic symptoms following artificial inoculation were water soaking of the infiltrated zone between 2 and 8 days after inoculation (dai); raised epidermis accompanying tiny yellow colored bacterial colonies at 8 dai; and yellowing and necrosis of the infected zone by 12–16 dai. In planta Xac population increased 1000 fold by 14 dai from an initial population of 7.3 × 10⁶ cfu cm⁻² (0 dai). Peak concentrations of H₂O₂ were observed at 24 h and between 8 and 10 dai and coincided with higher activity of total superoxide dismutase (SOD). Lower levels of H₂O₂ in infected leaves were maintained by Xac induced higher activities of catalase (CAT), ascorbate peroxidase (APOD), and guaiacol peroxidase (POD). It appears Xac altered H₂O₂ metabolism in C. sinensis L. Osb. ‘Hamlin’ to enhance survival and growth.     

Identification of S-alleles in pear (Pyrus communis L.) cv. ‘Rocha’ and other European cultivars

In this work we report the cloning and identification of S-RNase alleles responsible for gametophytic self-incompatibility (GSI) of ‘Rocha’ pear and of 13 other European pear cultivars that might be used as its pollinators. Partial sequences of S-RNase alleles were amplified by PCR with specific primers hybridising in conserved regions of previously identified S-RNase alleles of Pyrus communis, cloned and sequenced and the S-genotype of eight pear cultivars was fully determined. Three cultivars (‘General Léclerc’ (SqSl), ‘Tosca’ (SbSl) and ‘Alexandrine Douillard’ (SbSk)) shared no S-alleles with ‘Rocha’ (SaSe) and shall be totally compatible with this cultivar. None of the cultivars analysed showed an identical amplification pattern to the one observed in ‘Rocha’, so the other cultivars shall be at least semi-compatible. One new allele was identified in P. communis cv. ‘Beurré d’Avril’ (designated as St). The determination of both S-RNase alleles of cvs ‘Rocha’, ‘Beurré Precoce Morettini’ (SeSk) and ‘Tosca’ and the identification of one S-RNase allele in cvs ‘Carapinheira’ (Sb), ‘Amêndoa’ (Se), ‘Pérola’ (Sk) and ‘Beurré d’Avril’ (St) are important contributions for the effort recently developed worldwide to establish groups of sexual compatibility among European pears.     

Harvest and post-harvest apple fruit quality following applications of kaolin particle film in South African orchards

Sunburn on apples in the Western Cape region of South Africa can result in yield losses of up to 50%. Application of kaolin-based particle film (Surround^® WP) increases leaf and fruit surface reflectivity, thus potentially reducing heat load and sunburn. Trials were performed during 2002/2003 and 2003/2004 to assess the effectiveness of kaolin particle film in controlling sunburn on ‘Granny Smith’, ‘Braeburn’, ‘Fuji’, ‘Royal Gala’ and ‘Cripps’ Pink’ apples. Fruit quality and ripening were monitored following 1–4 months in regular atmosphere cold storage at −0.5 °C. Sunburn on exposed fruit was significantly reduced in ‘Granny Smith’ and ‘Fuji’. Kaolin treatment improved fruit colour of ‘Granny Smith’ and ‘Royal Gala’, and delayed starch conversion in ‘Granny Smith’ at harvest and during the early storage period but not thereafter. Incidence of watercore at harvest was significantly reduced by kaolin treatment, but this disorder disappeared during cold storage in both treatments. There were no effects on skin anthocyanin or phenolic concentrations in any cultivar compared to unsprayed fruit.