Influence of mitochondria origin on fruit quality in a citrus cybrid

Sugar, organic acid, and carotenoid are the most important indicators of fruit taste and nutritional and organoleptic quality. These components were studied on fruit pulp of the cybrid between Willow leaf mandarin ( Citrus deliciosa Ten.) and Eureka lemon [ Citrus limon (L.) Burm.] and the two parents. The cybrid possessed nuclear and chloroplast genomes of Eureka lemon plus mitochondria from Willow leaf mandarin. The impact of new mitochondria on fruit quality was studied during the mature period. Levels of organic acids were slightly higher in the cybrid fruit pulp than in Eureka lemon. No significant difference in sugar and carotenoid content was observed between the cybrid and the lemon. Results confirm that the main genetic information for the biosynthesis of sugars, organic acids, and carotenoids is contained in the nucleus. In Citrus, cybridization can be used as a strategy to breed specific traits associated with mitochondrial genomes, such as male sterility, without affecting the main organoleptic and nutritional qualities.     

Leaf volatile compounds of seven citrus somatic tetraploid hybrids sharing willow leaf mandarin (Citrus deliciosa Ten.) as their common parent

Volatile compounds were extracted by a pentane/ether (1:1) mixture from the leaves of seven citrus somatic tetraploid hybrids sharing mandarin as their common parent and having lime, Eurêka lemon, lac lemon, sweet orange, grapefruit, kumquat, or poncirus as the other parent. Extracts were examined by GC-MS and compared with those of their respective parents. All hybrids were like their mandarin parent, and unlike their nonmandarin parents, in being unable to synthesize monoterpene aldehydes and alcohols. The hybrids did retain the ability, although strongly reduced, of their nonmandarin parents to synthesize sesquiterpene hydrocarbons, alcohols, and aldehydes. These results suggest that complex forms of dominance in the mandarin genome determine the biosynthesis pathways of volatile compounds in tetraploid hybrids. A down-regulation of the biosynthesis of methyl N-methylanthranilate, a mandarin-specific compound, originates from the genomes of the nonmandarin parents. Statistical analyses showed that all of the hybrids were similar to their common mandarin parent in the relative composition of their volatile compounds.     

Predominant expression of diploid mandarin leaf proteome in two citrus mandarin-derived somatic allotetraploid hybrids

Fusion of citrus diploid parental protoplasts generates allotetraploid hybrids which do not retain their parental traits with regard to leaf aroma compound biosynthesis. The aim of this study was thus to examine hybrid leaf proteomes in comparison with their parents. Leaf soluble proteins from two citrus allotetraploid hybrids (mandarin + lime and mandarin + kumquat) and their diploid parents (mandarin, lime, and kumquat) were submitted to 2-D gel electrophoresis. Leaf proteome maps of the tetraploid hybrids were compared with those of their parents on the basis of the presence/absence of spots and of their spot relative volumes. The two allotetraploid hybrid maps were found closer to that of their mandarin parent than to those of their nonmandarin parents in terms of the presence/absence of spots as well as at a quantitative level. This approach has to be related to the already observed dominance of mandarin in allotetraploids with regard to volatile compound biosynthesis in leaves.     

Comparison of volatile concentrations in hand-squeezed juices of four different lemon varieties

Fresh juices of four Italian cultivars of lemons (Citrus limon Burm) have been analyzed by headspace solid phase microextraction coupled to gas chromatography and gas chromatography-mass spectrometry (GC-MS). The best results have been obtained with the 2 cm 50/30 microm divinylbenzene/carboxen on poly(dimethylsiloxane) fiber, using a homogenization time of 1 h at 40 degrees C and a sampling period of 30 min. A total of 35 volatile compounds have been identified by GC-MS, and their relative amounts have been calculated by adding internal standard to the samples. Differences in composition of lemon juices volatile components have been observed. Verdello Siracusano lemon juice has the highest amount of volatile compounds (50.28 mg/L), followed by Interdonato (8.39 mg/L), Primo Fiore Capo d'Orlando (5.75 mg/L), and Femminello Siracusano (2.62 mg/L) juices. Volatile compounds mainly consist of mono- and sesquiterpene hydrocarbons and oxygenated molecules (aldheydes, monoterpene alcohols, and monoterpene esters). Headspace solid phase microextraction coupled to a gas chromatograph equipped with a specific sulfur detector, a sulfur chemiluminescence detector, let us detect and quantify dimethyl sulfide compound at the microgram/liter level in lemon juices.     

Citrus rootstock and carbon dioxide enriched irrigation influence on seedling emergence,growth, and nutrient content 1

Seeds of Carrizo citrange (Citrus senensis (L.) Osb. X Poncirus trifolliata (L.), Cleopatra mandarin (C. reticulata Blanco), Sour orange (C. aurantium L.), and Rough lemon (C. Union (Buna f.) were sown in trays, irrigated without or with enriched Carbon dioxide (CO₂) (1,362 mg L^‐1) and evaluated for seedling emergence, growth, and nutrient contents. Rough lemon had a faster rate and higher percent emergence than the other rootstocks. Carrizo citrange had thicker stem diameters and taller seedlings than other rootstocks. Cleopatra mandarin had the smallest seedling shoot and root weights and larger shootrroot ratios than Rough lemon and Sour orange. Carrizo citrange and Cleopatra mandarin had higher leaf chlorophyll‐a and total chlorophyll content than Rough lemon or Sour orange. Carbon dioxide enriched irrigation had no effects on emergence or seedling growth variables except lower root weight. Lower media pH (6.90 versus 5.65), attributed to CO₂ enriched irrigation, may have adversely affected root growth as compared to shoot characteristics. Leaf nutrient contents generally differed between rootstocks but were not affected by CO₂ enriched water except for higher Zn and lower Mn contents. These results indicate that citrus seedling emergence, subsequent growth and leaf nutrient content differred between rootstocks but there are no beneficial effect from CO₂ enriched irrigation.     

Humic Acid–Like Substances Extracted from Compost Improve Fe Nutrition of Lemon Grown on Calcareous Soil: An Environmentally Safe Approach

The short-term effectiveness of soil-applied humic acid–like substances [naturally complexed or uncomplexed with iron (Fe)] extracted from composted poultry (HLSP) and sheep (HLSS) manures in preventing lime-induced Fe chlorosis in Eureka lemon (Citrus lemon L.) cuttings grafted on sour orange (Citrus aurantium L.) was investigated and compared with the commonly applied ethylenediamine di-2-hydroxyphenyl acetate ferric (FeEDDHA). Treatments were untreated plants and plants fertilized with FeEDDHA, HLSP, and HLSS injected into the soil at a total rate of 25 mg Fe per plant each. Chlorophyll concentration index (CCI) of the youngest fully expanded leaves, growth vigor, and leaf Fe concentration were measured. HLSP and HLSS, similar to FeEDDHA, resulted in significantly greater CCI values, growth vigor, and leaf Fe concentration compared to untreated plants. HLSP and HLSS offer potential alternative to FeEDDHA to prevent lime-induced Fe deficiency in Eureka lemon. Concentrates of HLSP and HLSS can be commercially prepared to offer an inexpensive and environmentally safe product.     

Influence of rootstocks on ‘Orlando’ tangelo leaf elemental concentration

Abstract

The influence of 10 rootstocks on leaf elemental concentrations and their relationship with yield, tree growth, leaf fresh weight and dry matter of ‘Orlando’ tangelo (C. parodisi Macf. x C. reticulata Blanco) was studied under the arid climate of southwestern Arizona. ‘Orlando’ tangelo trees on Carrizo citrange [(C sinensis (L.) Osbeck x Poncirus trifoliata (L.) Raf.], Yuma citrange (P. trifoliata x C. sinensis), Taiwanica (C. taiwanica) and Volkamer lemon (C. limon Burm f.) had larger canopy and higher yield than the trees on Savage citrange (P. trifoliata x C. sinensis), Ichang pummelo (C. ichangensis hyb.) and Palestine sweet lime (C. limettoides Tan.). Leaves of trees on Carrizo citrange were heavier but those on Savage citrange had higher percent dry matter than those on other rootstocks. Trees on macrophylla (Alemow) (C macrophylla Wester) had significantly higher leaf N and Mn while those on Carrizo, Yuma and Savage citranges had lower leaf N than the trees on other rootstocks. Trees on Volkamer lemon, rough lemon (C. jambhiri Lush), macrophylla and Palestine sweet lime had low leaf K but high leaf Mg. ‘Orlando’ trees on Carrizo citrange had the highest leaf Mg and Cu but the lowest leaf Mn. Considering mineral (elemental) uptake efficiency, tree growth and/or yield, Carrizo citrange, Volkamer lemon, Yuma citrange, rough lemon and Taiwanica are desirable for ‘Orlando’ tangelo under the arid conditions and sandy soils of the southwest United States. Macrophylla could only be used for a short term planting, while Savage citrange, Batangas mandarin (C. reticulata Blanco), Ichang pummelo and Palestine sweet lime are not recommended for planting under the arid conditions similar to those of this experiment.     

Effectiveness of Vivianite to Prevent Lime-Induced Iron Deficiency in Lemon Trees Grown on Highly Calcareous Soil

The short-term effectiveness of three application rates of vivianite [(Fe₃(PO₄)₂·8H₂O)] in preventing lime-induced iron (Fe) chlorosis in Eureka lemon (Citrus lemon L.) cuttings grafted on sour orange (Citrus aurantium L.) was investigated and compared with the commonly applied iron ethylenediaminedi(o-hydroxyphenylacetic) acid (FeEDDHA). Treatments were suspension of vivianite injected into the soil at three rates (0.5, 1.0, and 2.0 g kg^?1 soil), 417 mg FeEDDHA per plant, and untreated plants. Chlorophyll concentration index (CCI) of the youngest fully expanded leaves was estimated. Growth vigor and leaf Fe concentration were also measured. Vivianite, particularly at the greatest two rates, resulted in significantly greater growth vigor and leaf Fe concentration and exhibited greater CCI values compared to untreated plants similar to FeEDDHA. However, if excessive growth vigor is not favorable, the 0.5 g vivianite kg^?1 soil is recommended for farmers. Vivianite is a potential environmentally safe alternative to the expensive FeEDDHA to prevent Fe chlorosis in lemon.     

FOLIAR APPLICATION OF BORON INFLUENCES THE LEAF MINERAL STATUS,VEGETATIVE AND REPRODUCTIVE GROWTH,YIELD AND FRUIT QUALITY OF ‘KINNOW’ MANDARIN (CITRUS RETICULATA BLANCO.)

The effects of foliar application of boron (B) in the form of boric acid on vegetative and reproductive growth, yield and fruit quality of ‘Kinnow’ mandarin (Citrus reticulate Blanco.) were investigated. Kinnow mandarin was sprayed with different concentrations of boric acid viz. (0.1%, 0.2%, 0.3% and 0.4%) at fruit set stage. Leaf nitrogen (N), phosphorus (P), potassium (K), B, and zinc (Zn) along with flush length, tree height, tree spread and tree trunk diameter increased, while leaf length and leaf age showed non-significant results after foliar B application. Yield of the ‘Kinnow’ mandarin was significantly affected by foliar application of B and a significant increase in fruit weight at harvest was also observed. Soluble solid concentration (SSC): titratable acidity (TA) ratio, ascorbic acid, total sugars, total phenolic content (TPC) and total antioxidants significantly affected, while pH of juice, SSC, TA, reducing sugars and non-reducing sugars showed non- significant results.     

Involvement of limonene hydroperoxides formed after oil gland injury in the induction of defense response against Penicillium digitatum in lemon fruit

The effects of wounding oil glands of lemon [Citrus limon (L.) Burm.] fruit were investigated. Young mature-green lemons demonstrated significantly lower decay incidence than older yellow fruit when their oil glands were punctured in the presence of postharvest wound pathogen Penicillium digitatum Sacc. Contact with the released gland content on the green lemon surface reduced the viability of P. digitatum spores approximately twice. Wounding caused rapid production of limonene hydroperoxides that persisted for only a few minutes. The magnitude depended on the physiological maturity of the fruit; mature-green fruit produced much higher levels than did yellow lemons. Furthermore, wounding of the oil glands or injection of limonene hydroperoxides into the lemon peel elicited the production of the citrus fruit phytoalexins, scoparone and scopoletin, to levels known to be effective in reducing decay caused by P. digitatum. The mature-green fruit produced about twice as much of these phytoalexins as the older yellow fruit. This induced defensive elicitation of phytoalexin production, as well as the direct effects of these antifungal compounds, markedly inhibited the pathogen in mature-green fruits but was ineffective in older yellow ones.     

Simultaneous determination of Cd(II), Cu(II), Pb(II), and Zn(II) in citrus essential oils by derivative potentiometric stripping analysis

Citrus essential oils are widely used in the food, cosmetics, and pharmaceutical industries, so the determination of heavy metals content is of great importance to guarantee their quality. The present work deals with the quantification of Cd(II), Cu(II), Pb(II), and Zn(II) in different varieties of citrus essential oils, using derivative potentiometric stripping analysis. Two different metals extraction procedures, involving concentrated hydrochloric acid treatment and acid-alcoholic dissolution, are tested on lemon, mandarin, sweet orange, and bergamot essential oils, and they give very similar results. Cd(II), Cu(II), Pb(II), and Zn(II) recovery tests spanned from 95 to 100.50%, providing evidence that metals quantification remained unaffected by the cleanup steps of the two procedures. The repeatability of the hydrochloric acid extraction method, applied on different varieties of essential oils, is >95.00% for Cd(II), Cu(II), Pb(II), and Zn(II), whereas the repeatability of the acid-alcoholic dissolution method is >93.00% for Cu and Cd only in lemon oil. Detection limits obtained for the four analytes, using both procedures, ranged from 0.10 to 0.98 ng g(-)(1) in lemon, mandarin, sweet orange, and bergamot essential oils.     

Volatile constituents of wild citrus Mangshanyegan (Citrus nobilis Lauriro) peel oil

Volatiles of a wild mandarin, Mangshanyegan (Citrus nobilis Lauriro), were characterized by GC-MS, and their aroma active compounds were identified by aroma extract dilution analysis (AEDA) and gas chromatography-olfactometry (GC-O). The volatile profile of Mangshanyegan was compared with those of other four citrus species, Kaopan pummelo (Citrus grandis), Eureka lemon (Citrus limon), Huangyanbendizao tangerine (Citrus reticulata), and Seike navel orange (Citrus sinensis). Monoterpene hydrocarbons predominated in Mangshanyegan, in particular d-limonene and β-myrcene, which accounted for 85.75 and 10.89% of total volatiles, respectively. Among the 12 compounds with flavor dilution factors (FD) = 27, 8 oxygenated compounds, including (Z)- and (E)-linalool oxides, were present only in Mangshanyegan. The combined results of GC-O, quantitative analysis, odor activity values (OAVs), and omission tests revealed that β-myrcene and (Z)- and (E)-linalool oxides were the characteristic aroma compounds of Mangshanyegan, contributing to the balsamic and floral notes of its aroma.     

Juice components of a new pigmented citrus hybrid Citrus sinensis (L.) Osbeck x Citrus clementina Hort. ex Tan

Fruit juice of a new pigmented citrus hybrid named Omo-31 and those of its parents clementine cv. Oroval (Citrus clementina Hort. ex Tan.) and Moro orange [Citrus sinensis (L.) Osbeck] were analyzed during fruit maturation to determine juice yield, total soluble solids (TSS), total acidity (TA), TSS/TA ratio (classical parameters of quality), and potential health beneficial components, such as vitamin C, flavanones, anthocyanins, and phenolic acids. Results showed that juice yield, TA, TSS, and TSS/TA ratio values of Omo-31 were similar to those of the Moro orange. Vitamin C content of the new hybrid was slightly higher than that of clementine and lower than that of the Moro orange, but at maturity stage no differences were observed among the three genotypes. The phenolic compounds content of the new hybrid and those of the parents and their evolution during maturation were studied. At maturity stage the amount of anthocyanins, flavanones, and hydroxycinnamic acids in Omo-31 was found to be notably higher than those of the parents. The high level of antioxidant substances makes this new fruit important for its nutritional benefits.     

缺磷胁迫加重柑橘叶片光合作用的光抑制及叶黄素循环的作用

用营养液培养的方法 ,研究了不同光强和磷水平对温州蜜柑叶片光合作用光抑制的影响。结果表明 ,在缺磷强光条件下 ,叶片的初始荧光Fo、最大荧光Fm、PSⅡ光化学效率 (Fv/Fm)及表观电子传递速率 (ETR)下降 ,叶片非光化学猝灭的慢相 (qNs)和叶温升高。用叶黄素循环的抑制剂DTT处理叶片后 ,Fo升高。这些结果说明 ,缺磷胁迫加重了温州蜜柑叶片光合作用的光抑制 ,叶黄素循环的非辐射能量耗散在保护光合机构免受强光破坏方面起着重要作用。     

Estimating bergamot juice adulteration of lemon juice by high-performance liquid chromatography (HPLC) analysis of flavanone glycosides

The chemical composition of 30 samples of juices obtained from bergamot (Citrus bergamia Risso and Poit.) fruits is reported and compared to the genuineness parameters adopted by Association of the Industry of Juice and Nectars (AIJN) for lemon juice. It was found that the compositional differences between the two juices are distinguishable, although with difficulty. However, these differences are not strong enough to detect the fraudulent addition of bergamot juice to lemon juice. Instead, we found the high-performance liquid chromatography (HPLC) analysis of the flavanones naringin, neohesperidin, and neoeriocitrin, which are present in bergamot juice and practically absent in the lemon juice, is a convenient way to detect and quantify the fraudulent addition of bergamot juice. The method has been validated by calculating the detection and quantification limits according to Eurachem procedures. Employing neoeriocitrin (detection limit = 0.7 mg/L) and naringin (detection limit = 1 mg/L) as markers, it is possible to detect the addition of bergamot juice to lemon juice at the 1% level. When using neohesperidin as a marker (detection limit = 1 mg/L), the minimal percentage of detectable addition of bergamot juice was about 2%. Finally, it is reported that the pattern of flavonoid content of the bergamot juice is similar to those of chinotto (Citrus myrtifolia Raf) and bitter orange (Citrus aurantium L.) juices and that it is possible to distinguish the three kinds of juices by HPLC analysis.     

Salt tolerance of two lemon scions measured by leaf chloride and sodium accumulation

Effects of salinity on growth, ion content, water relationships, and chlorophyll and proline levels were measured on one‐year‐old ‘Verna’ and ‘Fino’ lemon (Citrus limon [L] Burm. F.) scions budded to either Sour orange (C. aurantium L.) or macrophylla (C. macrophylla Wester) rootstock. Trees were grown in nutrient solutions containing 2 (control), 40, or 80 mol m^‐3 NaCl for 75 days.

Growth of all combinations was reduced by salinity, but this effect was greater for both scions budded on macrophylla. Leaf chloride and sodium concentrations were lower in both scions budded on Sour orange. Leaf salt concentration was scion dependent. Leaves of ‘Fino’ lemon had higher levels of both chloride and sodium than did leaves of ‘Verna’ lemon, regardless of the rootstock considered.

Despite an accumulation of chloride and sodium in the leaves of salinized trees, leaf water potential and leaf water content increased above the control level. However, stomatal conductance declined in all rootstock/scion combinations.

Chlorophyll contents were markedly reduced by salt treatment; greater reductions were seen in ‘Fino’ lemon than in ‘Verna’ lemon on both rootstocks. Chlorophyll reductions were highly correlated with both chloride and sodium concentrations in the leaves. Free proline increased with salinity in leaves of both scions budded on Sour orange, but was unaffected on macrophylla.

Differences in the parameters determined in response to salinity were attributed to the different capacity of each specific rootstock/scion combination to import chloride and sodium into leaves.     

Contamination of Italian citrus essential oils: presence of chloroparaffin

In this study, the contamination by chloroparaffin of Sicilian and Calabrian citrus essential oils, produced in the crop years 1994-1996, was investigated. The analyses were carried out on 102 lemon oils, 98 orange oils, and 96 mandarin oils, using a dual-channel GC-ECD. It was found that 53% of lemon oil, 33% of orange oil, and 38% of mandarin oil samples were contaminated. The mean contamination levels were 7.1 ppm (lemon), 2.5 ppm (orange), and 5.3 ppm (mandarin). The highest concentration of chloroparaffin found was 60 ppm in a lemon oil sample.     

Leaf volatile compounds of six citrus somatic allotetraploid hybrids originating from various combinations of lime, lemon, citron, sweet orange, and grapefruit

Volatile compounds were extracted by a pentane/ether (1:1) mixture from the leaves of six citrus somatic allotetraploid hybrids resulting from various combinations of lime, lemon, citron, sweet orange, and grapefruit. Extracts were examined by gas chromatography-mass spectrometry (GC-MS) and compared with those of their respective parents. All hybrids having an acid citrus parent exhibit the same relative contents in hydrocarbons and oxygenated compounds as the acid citrus, while the (grapefruit + orange) hybrid behaves similarly to its two parents. When volatile compound contents (microg g(-1)) are examined in detail, several behaviors are encountered in hybrids and seem to depend on the presence/absence of the considered parental compound and on the corresponding hybrid combination. Meanwhile, the sesquiterpene hydrocarbons are present in all hybrids at concentrations systematically lower than those of the highest parental producers. Statistical analyses show that hybrids exhibit hardly discriminable aromatic profiles, meaning that no strong dominance of one or the other parent was observed in hybrids with regards to the leaf volatile compound production.     

Evaluation of auraptene content in citrus fruits and their products

Auraptene quantities in Tanaka's 77 Citrus species (including 14 varieties and cultivars), 5 Fortunella species, one Poncirus species, 27 hybrids between Citrus species, and 51 intergeneric hybrids between Citrus and Poncirus have been evaluated. The genus Citrus has been divided into eight groups. Auraptene is found in all of the species of Cephalocitrus group, a part of the species of Aurantium group, and most of the species of Osmocitrus group. The Citrus species contain a small amount of auraptene in the juice sacs compared with in the peels except for Henka mikan (C. pseudo-aurantium), Ichang lemon (C. wilsonii), and a Hassaku (C. hassaku)-pummelo hybrid (Okitsu No. 39), which contain large quantities of auraptene in their juice sacs (0.23, 0.52, and 0.14 mg/g, respectively). The Hong Kong wild kumquat (F. hindusii) alone contains auraptene in Fortunella species. All of the Citrus-trifoliate orange (P. trifoliata) hybrids as well as the trifoliate orange contain a large quantity of auraptene in both the peel (16.57-0.51 mg/g) and the juice sac (10.32-0.15 mg/g). These hybrids are almost inedible. The Iyo (C. iyo)-trifoliate orange hybrid (IyP269) is edible and contains auraptene in the peel (1.49 mg/g) and in the juice sac (1.73 mg/g). Citrus fruit products, for example, brand-named grapefruit juice and marmalade, retain about 0. 1 mg and 0.3 mg/100 g of auraptene, respectively.     

Betaines in fruits of Citrus genus plants

Numerous compounds, many of them osmolytes, were quantified in natural juices and in frozen concentrate juices from fruits of plants of the Citrus genus. L-proline, N-methyl-L-proline (hygric acid), N,N-dimethyl-L-proline (stachydrine), 4-hydroxy-L-prolinebetaine (betonicine), 4-hydroxy-L-proline, γ-aminobutyric acid (Gaba), 3-carboxypropyltrimethylammonium (GabaBet), N-methylnicotinic acid (trigonelline), and choline in the fruit juices of yellow orange, blood orange, lemon, mandarin, bitter orange (Citrus aurantium), chinotto (Citrus myrtifolia), and grapefruit were analyzed by sensitive HPLC-ESI-tandem mass spectrometry procedure. It was found that the most represented osmolytes in the juices, that is, L-proline, stachydrine, and betonicine, can be quantified with minimal sample preparation and short analysis time (about 1 min) also by flow injection analysis (FIA) ESI-MS/MS with the same results as obtained by HPLC ESI-MS/MS. In all of the juices, discrete amounts of choline and trigonelline were present. Conversely, GabaBet was always below detection limits. Notably, N-methyl-L-proline and 4-hydroxy-L-prolinebetaine, which were discovered for the first time in the juice of bergamot (Citrus bergamia Risso et Poit), are also present in all of the citrus juices examined.