Production of mandarin + pummelo somatic hybrid citrus rootstocks with potential for improved tolerance/resistance to sting nematode

Sting nematode (Belonolaimus longicaudatus Rau) has become a primary factor limiting citrus production in localized regions of the central Florida sandridge citrus production area, making the development of resistant rootstocks a new breeding objective. In efforts to develop a replacement rootstock for the widely adapted sour orange, our focus has been on somatic hybridization of selected mandarin + pummelo combinations [Grosser, J.W., Gmitter, Jr., F.G., 1990. Protoplast fusion and citrus improvement. Plant Breed. Rev. 8, 339–374; Ananthakrishnan, G., Calovic, M., Serrano, P., Grosser, J.W., 2006. Production of additional allotetraploid somatic hybrids combining mandarins and sweet oranges with pre-selected pummelos as potential candidates to replace sour orange rootstock. In Vitro Cell. Dev.: Plant 42, 367–371], since sour orange is probably an introgression hybrid of mandarin and pummelo as suggested by molecular marker analyses [Nicolosi, E., Deng, Z.N., Gentile, A., La Malfa, S., Tribulato, E., 2000. Citrus phylogeny and genetic origin of important species as investigated by molecular markers. Theor. Appl. Genet. 100, 1155–1166; Gulsen, O., Roose, M.L., 2001. Lemons: diversity and relationships with selected Citrus genotypes as measured with nuclear genome markers. J. Am. Soc. Hort. Sci. 126, 309–317]. Somatic hybrid plants were produced from four new mandarin (C. reticulata Blanco) + pummelo (C. grandis L. Osbeck) parental combinations by fusing embryogenic suspension culture-derived protoplasts isolated from selected mandarins with leaf protoplasts of pummelo seedlings previously selected for tolerance/resistance to the sting nematode (B. longicaudatus Rau) as follows: Amblycarpa mandarin + ‘Liang Ping Yau’ (seedling) pummelo seedling SN7; Amblycarpa mandarin + ‘Hirado Buntan Pink’ (HBP) pummelo seedling SN3; Murcott tangor + pummelo seedling SN3; and Shekwasha mandarin + pummelo seedling SN3. Somatic hybridization was verified by ploidy analysis (via flow cytometry) and RAPD analyses. Mandarin parents were selected for wide soil-adaptation and ability to produce friable embryogenic callus lines. Pummelo seedlings used as leaf parents were identified from a previous screen of large seed populations (200 each) from four pummelos for resistance to sting nematode as follows: ‘Hirado Buntan Pink‘; ‘Red Shaddock‘; ‘Large Pink Pummelo’ and a seedling pummelo of ‘Liang Ping Yau‘. Ten resistant/tolerant pummelo seedlings were selected from the 800 pummelo seeds planted in the screen for further study. The four new somatic hybrids have been propagated to evaluate their horticultural performance and resistance to the sting nematode. These potential somatic hybrid rootstocks should also have potential to control tree size due to polyploidy.     

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.     

‘Swingle’ citrumelo propagation by cuttings for citrus nursery tree production or inarching

‘Swingle’ citrumelo [Citrus paradisi MacFaden × Poncirus trifoliata (L.) Raf.] has been extensively used as a rootstock in several citrus growing regions of the World, including Southern Brazil where ‘Rangpur’ lime (Citrus limonia Osbeck) is still the predominant variety despite being affected by several important pathogens. In this case, ‘Swingle’ citrumelo is used to produce nursery trees to establish new orchards or to be inarched in adult and healthy groves in order to change the rootstock. We report herein a system to produce trees on ‘Swingle’ citrumelo more rapidly by budding onto non-rooted cuttings, as well as assessing potential to rapidly multiply ‘Swingle’ through rooting of non-budded cuttings. Therefore, two potential products are described: budded trees and rooted rootstock cuttings. ‘Valencia’ sweet orange [Citrus sinensis (L.) Osbeck] was budded at different heights on cuttings derived from eight-month old rootstocks. Grafted and additional non-budded cuttings were then treated with indole-3-butyric acid (500 mg L⁻¹) or left untreated before rooting. Three types of cuttings were evaluated: softwood, semi-hardwood and hardwood. The use of nursery trees derived from pre-budded hardwood cuttings of ‘Swingle’ citrumelo is an alternative grafting method on this cultivar. Softwood cuttings with one leaf pair were considered the most adequate material for rapid multiplication of ‘Swingle’ citrumelo by cutting. This could be particularly useful for inarching production or conventional budding after transplant of cutting-derived rootstocks.     

Horticultural performance of ‘Folha Murcha’ sweet orange onto twelve rootstocks

Despite its outstanding position, the Brazilian citriculture is established on a very limited pool of varieties that limits its expansion and restricts the fruit availability throughout the year. This situation determines the urgent necessity of developing alternative scion and rootstock cultivars, with good performance under local conditions. ‘Folha Murcha’ sweet orange (Citrus sinensis (L.) Osbeck) is a late-harvest cultivar, suitable both for the juice processing industry and the fresh fruit market, being described as tolerant to citrus canker (Xanthomonas citri subsp. citri Schaad et al.), and less affected by citrus variegated chlorosis (Xylella fastidiosa Wells et al.). A study was conducted in Bebedouro, São Paulo State, Brazil, to evaluate the horticultural performance of ‘Folha Murcha’ sweet orange 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 ‘Sun Chu Sha Kat’ mandarin (C. reticulata Blanco); the ‘Sunki’ mandarin (Citrus sunki (Hayata) Hort. ex. Tanaka); the ‘Rangpur’ limes (C. limonia Osbeck) ‘Cravo Limeira’ and ‘Cravo FCAV’; ‘Carrizo’ citrange (C. 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 7 m × 4 m spacing, in a randomized block design, with five replications and two plants per plot. No supplementary irrigation was applied. Fruit yield, canopy volume, tree tolerance to drought and to citrus variegated chlorosis, and fruit quality were assessed for each rootstock. Trees grafted onto the ‘Flying Dragon’ trifoliate were smaller in size, but had largest yield efficiency when compared to those grafted onto other rootstocks. Lower alternate bearing index was observed on trees budded onto ‘Cravo FCAV’ ‘Rangpur’ lime. Both ‘Rangpur’ lime rootstocks and the ‘Sunki’ mandarin induced higher tree tolerance to drought. The ‘Flying Dragon’ trifoliate induced better fruit quality and higher tolerance to citrus variegated chlorosis (CVC) to ‘Folha Murcha’ trees. A cluster multivariate analysis identified three groups of rootstocks with similar effects on ‘Folha Murcha’ tree performance. Among the 12 evaluated rootstocks, the ‘Flying Dragon’ trifoliate has a unique effect on plant growth, tolerance to drought and CVC, fruit yield and fruit quality of ‘Folha Murcha’ trees, and may be better suited for high-density plantings.     

Presence of diverse ratios of lycopene/β-carotene in five pink or red-fleshed citrus cultivars

Pink or red-fleshed fruit mutations are commonly found in grapefruit, sweet orange, and occasionally in lemon, which combine novel appearance with fine eating quality. In order to identify the major coloured pigments, high performance liquid chromatography (HPLC) with ultraviolet detection was applied for the separation and characterization of carotenoids from five pink or red-fleshed citrus cultivars. As a result, both lycopene and β-carotene with similar HPLC profiles were detected in the five citrus cultivars, ‘Fengdu’ red-fleshed pomelo (Citrus grandis (L.) Osbeck), ‘Guanxi’ sweet pomelo red mutant, ‘Hirado Buntan’ pomelo, ‘Cara Cara Navel’ orange (C. sinensis (L.) Osbeck) and ‘Star Ruby’ grapefruit (C. paradisi Macf.). ‘Star Ruby’ contained the highest lycopene and β-carotene content in its flesh. However, although the significant correlation between the concentrations of lycopene and β-carotene was detected with a coefficient of 0.9692 (P < 0.01), ratios of lycopene/β-carotene were different among the cultivars. Post-harvest biosynthesis verified that the flesh itself synthesized carotenoids rather than acquiring them via transport from other tissues. In addition, a product feedback regulation mechanism might be involved in the process of carotenoid biosynthesis.     

Cellular wall metabolism in citrus fruit pericarp and its relation to creasing fruit rate

Citrus creasing results in serious economic loss in many citrus orchards. Based on the different incidence of creasing, two sweet orange cultivars ‘Hong Jiang’ (grafting chimaera of which flesh is mostly from Citrus reticulata Blanco and peel is from Citrus sinensis Osbeck in) and ‘An Liu’ (Citrus sinensis Osbeck) were used to investigate the creasing rate. The ultrastructure in cellular wall, cellular wall component, cellular wall degradation-related enzymes and expansin (Ct-Exp1) of sweet orange during fruit ripening in field were comparatively analyzed. ‘Hong Jiang’ sweet orange had a higher creasing rate than ‘An Liu’ during ripening. The activities of polygalacturonase, cellulase and pectinesterase in cellular wall of ‘Hong Jiang’ increased more markedly compared with ‘An Liu’. The increases in the content of soluble pectin, ionically associated pectin, covalently bound pectin, hemicellulose and cellulose of ‘Hong Jiang’ were higher than those of ‘An Liu’. Furthermore, the enhanced degradation of the ultrastructure in the albedo cellular walls was observed in ‘Hong Jiang’. Moreover, the northern blot analysis indicated that the Ct-Exp1 gene expressed more strongly in peel of ‘Hong Jiang’ than ‘An Liu’ during fruit ripening. These data suggest that enhanced loss of pectin and cellulose in the cellular walls of peel tissue of sweet orange could result in fruit creasing.     

Evaluation of rootstocks for the Cyprus local lemon variety ‘Lapithkiotiki’

The effect of various rootstocks on yield, yield efficiency, tree size and fruit quality of the local lemon variety ‘Lapithkiotiki’ (Citrus limon (L.) Burm. F.) was studied under Cyprus conditions. Total cumulative yield over 13 years of production was the highest on rough lemon (C. jambhiri Lush) followed by that on Volkameriana (C. volkameriana Ten. & Pasq.), Yuma Ponderosa lemon (C. limon (L.) Burm. F.), sour orange (C. aurantium L.), Citrus macrophylla Wester, Morton citrange (C. sinensis cv. Washington navel × Poncirus trifoliata (L.) Raf.), Yuma citrange, Rangpur lime (C. limonia Obs.), Palestine sweet lime (C. limettioides Tan.), C-32 citrange and Citremon 1449 (C. limon × P. trifoliata), although no statistically significant differences were found between sour orange, the commercial rootstock used in Cyprus, and the other above mentioned rootstocks. Next in order as regards to total cumulative yield was Cleopatra mandarin (C. reticulata Blanko.) followed by Carrizo citrange, with statistically significant differences compared with sour orange. Trees on Citrumelo CPB-4475 (C. paradisi Macf. × P. trifoliata) and C-35 citrange died 3–4 years after grafting. Canopy volume was the lowest for trees on Cleopatra mandarin and Carrizo citrange. Yield efficiency A, expressed as total cumulative yield per trunk cross-sectional area, was the highest on C. macrophylla and lowest on Carrizo citrange. Rootstock significantly affected fruit size and weight, rind thickness, juice content, brix, total acids and brix:acid ratio. The results of the present study reveal that the most promising rootstocks that may replace sour orange for the local lemon variety ‘Lapithkiotiki’ under Cyprus conditions are Volkameriana, Yuma Ponderosa lemon, C. macrophylla and Citremon 1449.     

Agrobacterium tumefaciens-mediated genetic transformation and plant regeneration from a complex tetraploid hybrid citrus rootstock

Agrobacterium-mediated genetic transformation of a tetraploid “tetrazyg” citrus rootstock selection ‘Orange #16’ [Nova mandarin (Citrus reticulata Blanco) + Hirado Buntan pummelo (Citrus grandis L. Osbeck)] × [Cleopatra mandarin (C. reticulata Blanco) + Argentine trifoliate orange (Poncirus trifoliata (L.) Raf.)] was performed. Juvenile epicotyl segments were transformed with a construct containing a bifunctional egfp–nptII fusion gene under the control of an enhanced double CaMV 35S promoter. Our protocol resulted in a reasonable transformation efficiency of 18%. Stable integration of the transgene was confirmed by visual observation of EGFP expression, PCR and Southern blot hybridization. The purpose of this work was to investigate the amenability of novel citrus rootstock germplasm being developed for improved tree size control, soil adaptation, and disease resistance, to existing transformation technologies. Seed trees of such transgenic tetraploids also have potential as trap plants containing potent insecticidal transgenes, due to their inedible fruit and inherent crossing barriers with conventional commercial diploid scion cultivars, and could be planted around producing citrus groves.     

Characteristics and oleocellosis sensitivity of citrus fruits

The effects of variety, growth phase, and water loss on development of oleocellosis, and relationships between chromatism and Vis/NIR spectra were studied in ‘EarlyGold’ sweet orange (Citrus sinensis Osbeck), ‘Fukumoto’ navel (Citrus sinensis Osbeck), and ‘Cara Cara’ navel (Citrus sinensis Osbeck) oranges. The varieties showed significant differences in the rate (RO) and degree (DO) of oleocellosis development. The sensitivity of varieties (from most to least sensitive) was ‘EarlyGold’ > ‘Fukumoto’ > ‘Cara Cara.’ Growth phase and water loss had a significant influence on fruit sensitivity to oleocellosis. The order of sensitivity to oleocellosis was dependent on harvest time (i.e., at normal period > at delayed period > at uncolored period), and RO and DO decreased significantly with water loss. The RO and DO models for fruit water loss were established as y = 0.75 − 3.94x − 271.33x² (R² = 0.77) and y = 1.70 − 7.29x − 1025.83x² (R² = 0.583). The sensitivity of ‘EarlyGold’ to oleocellosis was significantly correlated with dL and dC of fruit chromatism. At the same time, there were significant differences at 480–575 nm, 650–720 nm, and 925–965 nm between varieties with low and high sensitivity to oleocellosis, and ‘EarlyGolds’ with a low RO and DO had a higher reflectance than those with a high RO and DO.     

柑桔体细胞杂种扦插繁殖及其有关性状研究

对３个由原生质体融合产生的柑桔体细胞杂种繁殖特性的研究表明，金柑与甜橙属间体细胞杂种扦插生根成活率较低。甜橙＋宜昌橙以及甜橙＋粗柠檬种间体细胞杂种扦插萌发和生根能力与权及粗柠檬没有显著差异；插条发根平均为２．０条和１．８条，一年生植株根系总长分别为１２２．２ｃｍ和８０．７ｃｍ，与积和粗柠檬均无显著差异；粗柠檬＋甜橙杂种的须根数为３９条，株－１，与权（５４．５条·株－１）差异显著，而宜昌橙＋甜橙种间杂种（４７．７条·株－１）与积差异不显著；体细胞杂种的抗寒性有介于双亲之间的趋势     

Cybrid/hybrid plants regenerated from somatic fusions between male sterile Satsuma mandarin and seedy tangelos

Somatic hybridization provides an alternative for transferring mitochondria-encoded cytoplasmic male sterility (CMS). Herein, symmetric protoplast electrofusion was conducted between embryogenic callus protoplasts of Citrus unshiu Marc. cv. Guoqing No. 1 (G1), a CMS cultivar, and mesophyll-derived protoplasts of seedy ‘Page’ tangelo [C. reticulata Blanco × (C. reticulata Blanco × C. paradisi Macf.)] or ‘Nova’ tangelo [C. reticulata Blanco × (C. reticulata Blanco × C. paradisi Macf.)], to transfer CMS trait. Flow cytometry analysis showed that 14 plants recovered from G1 + ‘Page’ tangelo that displayed typical morphological character of ‘Page’ were diploid, and 6 plants regenerated from G1 + ‘Nova’ tangelo were tetraploid. Genetic compositions of regenerated plants from the two fusions were determined by SSR, CAPS and chloroplast-SSR analysis. Cybrid nature of diploids from G1 + ‘Page’ tangelo with nuclear DNA from ‘Page’, mitochondrial DNA (mtDNA) from the G1 and chloroplast DNA (cpDNA) derived from either parent was confirmed. Tetraploid plants from G1 + ‘Nova’ tangelo were identified as somatic hybrids with random cpDNA inheritance. The regenerated cybrid and hybrid plants hold great potential for Citrus seedless breeding at diploid or triploid levels.     

Nitrogen best management practice for citrus trees: I. Fruit yield,quality, and leaf nutritional status

Elevated levels of nitrate-nitrogen (NO₃-N) in the surficial aquifer above the drinking water quality standard, i.e. maximum contaminant limit (MCL; 10 mg L⁻¹), have been reported in some part of central Florida citrus production regions. Soils in this region are very sandy (sand content >95%), hence are vulnerable to leaching of soluble nutrients and chemicals below the rooting depth of the trees. The objective of this research was to develop N and irrigation best management practices for citrus in sandy soils to maintain optimal crop yield and quality, and to minimize potential leaching of nitrate below the root zone. Six years of field experiment was conducted in a high productive (mean fruit yield > 80 Mg ha⁻¹yr⁻¹) >20-year-old ‘Hamlin’ orange trees [Citrus sinensis (L.) Osbeck] on ‘Cleopatra mandarin’ (Citrus reticulata Blanco) rootstock grown on a well drained Tavares fine sand (hyperthermic, uncoated, Typic Quartzipsamments) in Highland county, FL. Nitrogen rates ranged from 112 to 280 kg ha⁻¹ yr⁻¹ applied as fertigation (FRT), water soluble granular (WSG), 50:50 mix of FRT and WSG, and controlled-release fertilizer (CRF). Tensiometers were used to monitor the soil water content as a basis to schedule optimal irrigation. Fruit yield response over the entire range of N rates was greater for the FRT and WSG sources as compared to that for the WSG + FRT or CRF sources. Using the regression analysis of the fruit yield in relation to N rate, the optimum N rate appeared to be at 260 kg ha⁻¹ yr⁻¹. Based on fruit production response in this study, the N requirement for production of 1 Mg of fruit varied from 2.2 to 2.6 kg across four N sources. This study demonstrated an increased N uptake efficiency, as a result of best management of N and irrigation applications. The optimal N and K concentration in the 4–6-month-old spring flush leaves were 26–30, and 15–18 g kg⁻¹, respectively. However, fruit yield response showed no significant relationship with concentrations of P in the 4–6-month-old spring flush leaves over a range of 0.8–2.4 g kg⁻¹. The results of fate and transport of N in soil and in soil solution with application of different rates and sources of N, and components of citrus tree N budget, are reported in a companion paper.     

Plant growth,yield, and fruit quality of ‘Fallglo’ and ‘Sunburst’ mandarins on four rootstocks

Vegetative growth, yield, and fruit quality of ‘Fallglo’ and ‘Sunburst’ mandarins on ‘Rangpur’ lime, ‘Swingle’ citrumelo, ‘Orlando’ tangelo, and ‘Cleopatra’ mandarin were evaluated under subtropical climate of Northern São Paulo State, Brazil, from 2000 through 2006 harvest seasons. ‘Fallglo’ mandarin trees had the highest cumulative yield on ‘Rangpur’ lime, and the smallest on ‘Swingle’ citrumelo and ‘Orlando’ tangelo. Plants of this cultivar had the highest yield efficiency on ‘Rangpur’ lime, and the lowest on ‘Orlando’ tangelo. ‘Sunburst’ mandarin trees began to bear fruits later than ‘Fallglo’ mandarin trees, with no differences in yield induced by the rootstocks. ‘Cleopatra’ mandarin induced the most vigorous growth in ‘Fallglo’ mandarin as compared to plants on ‘Swingle’ citrumelo. On the other hand, the largest trees of ‘Sunburst’ mandarin were registered on ‘Orlando’ tangelo, and the smallest on ‘Rangpur’ lime. ‘Sunburst’ mandarin had higher alternate bearing than ‘Fallglo’ regardless the rootstock. Fruit weight and juice content were not affected by the rootstock. These two mandarin scion cultivars may be considered adequate alternatives to produce good fruit quality for the fresh fruit market. ‘Cleopatra’ mandarin and ‘Rangpur lime are suitable rootstocks for ‘Fallglo’ mandarin, whereas all rootstocks evaluated are adequate for ‘Sunburst’ mandarin.     

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.     

Chloride and sodium excluding capacities of citrus rootstock germplasm introduced to Australia from the People's Republic of China

The responses to root zone salinity (0, 25 and 50 mM NaCl) by 40 citrus rootstock genotypes introduced from the People's Republic of China, measured as shoot chloride (Cl⁻) and sodium (Na⁺) ion accumulation, growth and dry matter accumulation, were investigated under glasshouse conditions. Two experiments, one using aerated nutrient solutions and the other irrigated sand cultures, were conducted with plants grown from rooted cuttings taken from representative trees of different mandarin (Citrus reticulata Blanco and C. erythrosa Hort. ex Tan.), yuzu (C. junos Sieb. ex Tan.), Ichang papeda (C. ichangensis Swing.), sour orange (C. aurantium L.) and trifoliate orange (Poncirus trifoliata (L.) Raf.) genotypes. Two standard genotypes, viz. Rangpur lime (Citrus x limonia Osbeck.) and William's trifoliate orange were included in the experiments as reference controls.     

Nitrogen best management practice for citrus trees: II. Nitrogen fate,transport, and components of N budget

Soils in central Florida citrus production region are very sandy, hence are vulnerable to leaching of soluble nutrients and chemicals. The objective of this study was to develop nitrogen (N) and irrigation best management practices for citrus in sandy soils to maintain optimal crop yield and quality, and to minimize N leaching below the rootzone. A replicated plot experiment was conducted in a highly productive 20+ year-old ‘Hamlin’ orange [Citrus sinensis (L.) Osbeck] trees on ‘Cleopatra mandarin’ [(Citrus reticulata Blanco)] rootstock grove located on a well drained Tavares fine sand (hyperthermic, uncoated, Typic Quartzipsamments) in Highland County, FL. Nitrogen rates (112–280 kg ha⁻¹ year⁻¹) were applied as fertigation (FRT), water soluble granular (WSG), a combination of 50% FRT and 50% WSG, and controlled release fertilizer (CRF). Tensiometers were used to monitor the soil moisture content at various depths in the soil profile as basis to optimize irrigation scheduling. Fruit yield and quality and nutritional status of the trees were reported in a companion paper. Soil solution was sampled at 60, 120, and 240 cm depths under the tree canopy using suction lysimeters. Concentrations of NO₃-N in the soil solution at 240 cm deep, which is an indicator of NO₃-N leaching below the tree rootzone, generally remained below the maximum contaminant limit (MCL) for drinking water quality (10 mg L⁻¹) in most samples across all N sources and rates, but for few exceptions. Total N in the fruit was strongly correlated with fruit load, thus, at a given N rate N removal by the fruit was lower during years of low fruit yield as compared to that during the years of high fruit yield. Furthermore, there was a strong linear relation between N and K in the fruit. This supports the need to maintain 1:1 ratio between the rates of N and K applications. In a high fruit production condition, the N in the fruit accounted for about 45% of the total N input on an annual basis. Fifteen percent of the total N input at 280 kg N ha⁻¹ year⁻¹ was not accounted for in the citrus N budget, which could be due to leaching loss. This estimate of potential leaching was very close to that predicted by LEACHM simulation model. The improved N and irrigation management practices developed in this study contributed to an improved N uptake efficiency and a reduction in N losses.     

Performance of ‘Tahiti’ lime on twelve rootstocks under irrigated and non-irrigated conditions

Faced with new challenges, such as emerging diseases, shortening of orchard longevity, and larger social and environmental demands from consumers, practices such as rootstock diversification, irrigation and high density plantings have become relevant for the Brazilian citrus industry. This research had the objective to evaluate the performance of irrigated and non-irrigated ‘Tahiti’ lime trees grafted on 12 rootstocks and one interstock. Plots were distributed following a randomized block design, with four replicates and one plant per plot. Rootstocks influenced plant vigor, especially ‘Flying Dragon’ trifoliate, which reduced tree height by approximately 47% compared to the ‘Rangpur’ lime. Trees that were budded on more vigorous rootstocks showed higher yield when grown without irrigation than with irrigation. The ‘1646’ citradia and ‘Morton’ citrange rootstocks performed particularly well. On the other hand, the plants on less vigorous rootstocks showed better performance in terms of yield under irrigation than the same combinations without irrigation, especially those grafted on the tetraploid ‘Carrizo’ and ‘Troyer’ citranges, ‘Swingle’ citrumelo, ‘Davis A’ trifoliate and ‘Flying Dragon’ trifoliate. Plants budded on the ‘1708’ citradia had high yields under irrigated and non-irrigated conditions. The effect of interstock on plant vigor was dependent of rootstock. Interstocked plants on ‘Davis A’ trifoliate were higher than those without interstock. On the other hand, interstocked plants on Catania 2 ‘Volkamer’ lemon were less vigorous than those without interstock.     

Somatic hybridization of high yield,cold-hardy and disease resistant parents for citrus rootstock improvement

Summary

Increasing losses of trees to diseases, expansion of plantings into more marginal production areas and the need to control tree size to reduce harvesting costs have elevated the demand for new improved citrus rootstocks in Florida. A major strategy of the CREC rootstock improvement programme has been to use protoplast fusion to produce allotetraploid somatic hybrids that combine complementary rootstock germplasm. Tetraploid citrus rootstocks have been shown to have a built-in tree size control component due to polyploidy. This report focuses on the incorporation of newly identified superior rootstock germplasm into the somatic hybridization programme. Poncirus trifoliata 50–7, a selection of trifoliate orange selected for superior resistance to Phytophthora nicotianae, was hybridized with sour orange, Changsha mandarin, Navel orange, and a seedy white ‘Duncan’ type grapefruit. High-yielding Benton citrange (Citrus sinensis 3 Poncirus trifoliata) was hybridized with Changsha mandarin and sour orange. More than 200 plants of each of these six new somatic hybrids were propagated via tissue culture and rooted cuttings. To assess their horticultural performance, all the somatic hybrids were budded with commercially important scions and planted in replicated trials representing the two most important soil types in Florida. Seed trees of each somatic hybrid have also been planted in the field to determine fruiting potential and level of polyembryony.     

脐橙与粗柠檬体细胞杂种果实类胡萝卜素、糖酸遗传的亲本偏向性

 采用高效液相色谱测定柑橘种间体细胞杂种[‘朋娜’脐橙（Citrus sinensis Osbeck）+ 粗柠檬（C. jambhiri Lush）]和其两个融合亲本果实的类胡萝卜素含量，气相色谱测定可溶性糖及有机酸含量，并检测代谢过程中关键基因的表达，比较分析体细胞杂种果实品质遗传及基因表达特点。结果表明，体细胞杂种类胡萝卜素成分及含量均偏向粗柠檬亲本，柠檬酸、苹果酸积累量也偏向粗柠檬；而蔗糖含量处在中亲值；实时定量PCR 技术检测类胡萝卜素代谢途径中的7 个基因，其中4 个基因在‘朋娜’脐橙中的表达量高于粗柠檬，环化途径中番茄红素ε 环化酶基因（CitLcy-e）、玉米黄质环氧酶基因（CitZep）在体细胞杂种中表达量偏向粗柠檬亲本，显著低于‘朋娜’脐橙。可见粗柠檬遗传物质的表达在体细胞杂种中占主导地位。     

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.