important ( Ali et al., 2000 ). A species of multiple use, the sour orange ( Citrus aurantium L.) is also known as bitter or seville orange. It is a universal rootstock for citrus and is used widely in the Mediterranean region ( Navarro et al., 1975 ) It is
Kim D. Bowman and Frederick G. Gmitter Jr.
Chinotto is a selection of sour orange (Citrus aurantium L.) with short internodes and small leaves and fruit. Mature fruiting trees of Chinotto grafted on standard rootstocks produce healthy, but dwarf, trees. Most seedlings recovered from Chinotto fruit are of nucellar (maternal) origin and will faithfully grow to reproduce the Chinotto genotype and phenotype. Vigorous greenhouse-grown nucellar seedlings of Chinotto have internodes 5 to 10 mm in length and leaves 30 to 40 mm in length, about 30% the dimensions of the corresponding organs on standard sour orange nucellar seedlings. Sexual hybrids with Chinotto have been produced by controlled crosses with several other parents. Some hybrids with shortened internodes and small leaves were recovered among all hybrid progenies, regardless of whether Chinotto was used as seed or pollen parent. In some cases, segregation among Chinotto hybrids was about 1 normal: 1 dwarf. In other progenies, some intermediate forms were recovered along with normal and dwarf plants.
Sour orange seedlings were grown in water culture to which one of seven aromatic compounds, associated with allelopathic effects, was added to produce concentrations ranging from 0.5 to 2.0 mM. Leaf water potential (ψ1), leaf stomatal conductance (gs), and whole plant transpiration (T) were measured during a 7-day treatment period. At the end of that period, the total and average leaf surface area, shoot elongation, and fresh weight gain of seedlings were determined. Solutions of vanillic, coumaric, and ferulic acids of 2mM concentration reduced ψ1, gs, and T. Reductions of gs, and T but not (ψ1) occurred when vanillic acid of 1mM concentration was applied. Solutions of vanillic (0.5; 1.0; 2.0mM), coumaric (1; 2mM), cinnamic (1mM), or chlorogenic (1; 2mM) acids reduced fresh weight gain of seedlings. Only the coumaric and chlorogenic acids treatments of 2mM concentration reduced shoot elongation. No treatment affected total or individual leaf area. Gallic and caffeic acids had no effect on sour orange water relations and growth.
Amit Vikram, G.K. Jayaprakasha, and Bhimanagouda S. Patil
Our recent studies have shown that certain citrus limonoids protect from colon cancer based on cell and animal studies. Animal studies also suggest that citrus juice protects from osteoporosis. To understand the structure–function relationship through animal studies requires a large amount of purified limonoids. Since certain limonoids are present in low concentration, it is a challenge to obtain the required quantity of different limonoids. In this context, we report the purification of limonin 17-ß-D glucopyranosides (LG), and deacetylnomilinic acid 17-ß-D glucopyranoside (DNAG). However, DNAG was isolated in relatively large amount from sour orange (Citrus aurantium L.) seeds. Defatted seed powder was extracted with methanol and purified using column chromatography to obtain multigrams of DNAG. While LG was found in lower concentration, a large concentration of hesperidin was also purified in this process. This project is based upon work supported by the USDA-CSREES under Agreement USDA IFAFS #2001 52102 02294 and USDA #2005-34402-14401 “Designing Foods for Health” through the Vegetable & Fruit Improvement Center.
Jude W. Grosser, Victor Medina-Urrutia, Govindarajulu Ananthakrishnan, and Patricia Serrano
Sour orange has been a premier citrus rootstock worldwide due to its ability to perform on challenging soils and to produce and hold high-quality fruit. However, increasingly widespread quick-decline isolates of citrus tristeza virus (CTV) have destroyed entire industries on sour orange in some countries, and are in the process of destroying millions of trees on sour orange in Florida. CTV also threatens other citrus locations planted heavy to sour orange, including Texas and Mexico. An acceptable alternative rootstock to replace sour orange is in high demand but has yet to be developed. Molecular analyses have recently determined that sour orange is probably a hybrid of pummelo and mandarin. We report the production of 12 new mandarin + pummelo somatic hybrids produced by protoplast fusion from selected superior mandarin and pummelo parents, in efforts to develop a suitable replacement sour-orange-like rootstock that is resistant to CTV-induced quick decline. Somatic hybrids from all 12 parental combinations were confirmed by a combination of leaf morphology, flow cytometry, and RAPD analyses (for nuclear hybridity). These new mandarin + pummelo somatic hybrids are being propagated by rooted cuttings as necessary to conduct quick-decline resistance assays and to assess horticultural performance in replicated field trials.
Mirko Siragusa, Fabio De Pasquale, Loredana Abbate, and Nicasio Tusa
A collection of 18 accessions of sour orange (Citrus aurantium L.) coming from Sicily and other countries was investigated by two polymerase chain reaction (PCR)-based DNA marker technologies. Ten inter-simple sequence repeat (ISSR) primers and fifteen randomly amplified polymorphic DNA (RAPD) primers were used to identify and to evaluate the genetic variability and relationship of accessions. A total of 111 ISSR and 145 RAPD amplified fragments were used to estimate the Dice's coefficient of similarity for cluster analysis using a unweighted pair-group method using an arithmetic averaging (UPGMA) algorithm. The genetic relationships identified using ISSR and RAPD markers were highly concordant, such that the correlation between ISSR and RAPD genetic distance (GD) estimates was r = 0.93. The ISSR and RAPD analysis of 18 sour orange accessions found a high grade of genetic diversity in foreign accessions, while a low variability was detected in local accessions. Sicilian accessions could be grouped in two distinct clusters, including indistinctly plants from three origin regions. Some markers could be linked to the different growing areas. The ISSR and RAPD molecular reference system seems to be suitable for a fine identification of tightly related plants and the obtained results can form the basis for future setting up of Citrus rootstock genetic improvement projects.
David M. Eissenstat, James H. Graham, James P. Syvertsen, and Diana L. Drouillard
The effects of phosphorus (P) and of the mycorrhizal (M) fungus, Glomus intraradix, on the carbon (C) economy of sour orange (citrus aurantium L.) were determined during and following active M colonization. There were four treatments: mycorrhizal seedlings grown at standard-strength P (M1) and nonmycorrhizal (NM) plants grown at 1, 2 and 5 times standard-strength P (NM1, NM2 and NM5). Mycorrhizal colonization, tissue dry mass, P content, root length, leaf area, 14C partitioning and rate of c assimilation (A) were determined in five whole-plant harvests from 6 to 15 wks of age. In contrast to the effects of P nutrition on C economy in sour orange, M effects were generally subtle. Mycorrhizae increased the root biomass fraction, the root length/leaf area ratio, and the percent of 14C recovered from belowground components. Mycorrhizal plants had a higher percentage of belowground 14C in the respiration and soil fractions than did NM plants of equivalent P status. Mycorrhizal plants tended to have enhanced A at 8 wks but not at 7 or 12 wks. This temporarily enhanced A of M plants did not fully compensate for their greater belowground C expenditure, as suggested by apparently lower relative growth rates of M than NM plants of equivalent P status. Problems of interpreting the dynamic effects of mycorrhizae on C economy that are independent of p nutrition are discussed.
The purpose of this study was to determine the effect of Ca: NH4 ratio in the rhizosphere of hydroponically grown sour orange seedlings (SO) (Citrus aurantium L.) on the plants' vegetative growth and N uptake. The experiment was prompted by our observation that application of N in the form of NH4 in conjunction with CaCl2 was more efficient in eliminating N deficiency in field-grown grapefruit trees than the same rates of N applied in the form of NH4NO3 without CaCl2. About 40-cm-tall SO were pruned back to the 4th leaf and grown for 6 weeks in nutrient solutions containing 5 mm NH4 + at CaCl2: NH4 + molar ratios of 1.0, 1.3, 1.6, 1.9, 2.2, or 2.5. In an additional treatment, NO3 – was used as the sole source of N at CaCl2: NO3 – ratio of 1:1. The level of Ca:NH4 ratio had no effect on new leaves number, shoot growth, total and average leaf area, specific leaf weight, as well as leaf, stem, and tap root dry weight. However, lateral root dry weigh decreased at Ca: NH4 ratio of 2.5. No growth differences were found when the plants were supplied with NH4 + vs. NO3 – at Ca:N molar ratio of 1:1.
John E. Fucik and Dariusz Swietlik
Water extracts of cocklebur,CBX (Xanthium spinosa L.) and velvetleaf,VLX (Abutilon theophrasti Medic.) shoots and Mexican ash,AshX (Fraxinus Berlandieriana A.DC.) roots were added to 9 month-old sour orange Citrus aurantium L.) seedlings(SOs) in water culture. Final extract concentrations represented either 50 or 12.5 g. of plant material liter-1 of culture solution, i.e. 1/20 or 1/80 dilutions. Leaf water potential(ψ); stomatal conductance(gs);transpiration(T) and growth responses were measured for 13 days. After 1 day, SOs in AshX and CBX had lower ψ than controls. After 11 days SOs in CBX had higher ψ than the others. ψ responded similarly to both extract concs.. Thru day 5, AshX decreased gs vs. the controls and VLX. By day 11, gs of SOs in AshX was less than for VLX but not the others. On days 1 and 5, gs for VLX at 1/20 was lower than controls but at 1/80, gs's were the highest of all treatments. These results supported by the T rates, growth responses and others findings suggest AshX and VLX induce water stress by reducing water absorption and/or transport in addition to possibly disrupting normal root/shoot communications
Dariusz Swietlik and Linsen Zhang
Chelator-buffered nutrient solutions were used to study the effect of different levels of Zn activity in the rhizosphere on growth and nutritive responses of various tissues of sour orange seedlings. The seedlings were grown for 3 months in a growth chamber in a hydroponic culture containing from 5 to 69 μm and 5 to 101 μm total Zn in Expts. 1 and 2, respectively. Zn+2 activities were calculated with a computerized chemical equilibrium model (Geochem-PC), and buffered by inclusion of a chelator, diethylenetriamine pentaacetate (DTPA), at 74 and 44 μm in excess of the sum of Fe, Mn, Zn, Cu, Ni, and Co in Expts. 1 and 2, respectively. The use of DTPA-buffered solutions proved successful in imposing varying degrees of Zn deficiency. The deficiency was confirmed by leaf symptomatology, leaf chemical analyses, i.e., <16 mg·kg-1 Zn, and responses to foliar sprays and application of Zn to the roots. Growth parameters varied in their sensitivity to Zn deficiency, i.e., root dry weight < leaf number and white root growth < stem dry weight < leaf dry weight < shoot elongation and leaf area. The critical activities, expressed as pZn = -log(Zn+2), were ≈10.2±0.2 for root dry weight, 10.1±0.2 for leaf number and white root growth, 10.0±0.2 for stem dry weight, 9.9±0.2 for leaf dry weight, and 9.8±0.2 for shoot growth and leaf area. Increases in growth were observed in response to Zn applications even in the absence of visible Zn-deficiency symptoms. Seedlings containing >23 mg·kg-1 Zn in leaves did not respond to further additions of Zn to the nutrient solution. Zinc foliar sprays were less effective than Zn applications to the roots in alleviating severe Zn deficiency because foliar-absorbed Zn was not translocated from the top to the roots and thus could not correct Zn deficiency in the roots.