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Jinq-Tian Ling, N. Nito, and M. Iwamasa

Protoplasts were isolated from embryogenic calli of Citrus reticulata cv. Ponkan and Citropsis gabunensis, and fused in electric fields. The maximal fusion efficiency was obtained by application of AC at 75 V/cm (1.0 MHz) and DC square-wave pulse at 1.125 KV/cm for 40 usec. Fusion-treated protoplasts were cultured on MT medium without phytohormone, solidified with 0.6% agar. Colonies from the protoplasts were proliferated on MT medium with zeatin 1 mg/l and 0.9% agar. Selection of somatic hybrid callus was based on chromosome count and isoenzyme analysis. The somatic hybrids were tetraploid (2n=36). C. reticulata and C. gabunensis were both homozygous at Got-1 locus, but distinguishable easily because band of the latter migrated faster than that of the former. In zymogram of somatic hybrid, both parent bands were retained and a new hybrid band was also evident between them. Embryos from somatic-hybrid callus regenerated intact plant. The hybrid plants showed intermediate morphological characteristics of the parents.

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S. Singh, B.K. Ray, S. Bhattacharyya, and P.C. Deka

Multiple shoots were obtained from shoot tips (2 to 3 mm) derived from mature plants (5 to 6 years old) of Citrus reticulata Blanco cv. Khasi mandarin and C. limon Burm.f. cv. Assam lemon when cultured on Murashige and Skoog (MS) medium, supplemented with (mg·liter-1) 1.0 BAP, 0.5 kinetin, and 0.5 NAA. Root induction was observed when 7-week-old single shoots (≈ 2 cm long) of both Citrus species were cultured on MS medium supplemented with (mg·liter-1) 0.25 BAP, 0.5 NAA, and 0.5 IBA. These plantlets were successfully established in the soil. Chemical names used: naphthalene acetic acid (NAA), indole 3-butyric acid (IBA), and benzylamino purine (BAP).

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Mebelo Mataa and Shigeto Tominaga

The effects of root restriction, induced by root restriction bags, was evaluated on `Yoshida' Ponkan mandarin (Citrus reticulata Blanco). Trees were planted in 0.02-m3 volume root wrap bags (RWBs), which were made from woven polystyrene fiber, or root control bags (RCBs) made from nonwoven UV-stabilized Duon polystyrene fibre with plastic bottoms. A direct soil planted, nonrestricted root treatment (DPC) was included as a control. After 3 years, reductions in height (14% to 29%), canopy volume (66% to 43%), girth (10% to 22%), and leaf area (8% to 12%) were recorded in both of the root restriction treatments. Greater reductions occurred in the RWB treatment. Photosynthesis, transpiration, water potential, and leaf carbohydrate content were not affected by root restriction although soil moisture content was lower in the root restricted treatments. Fruiting efficiency (i.e., number of fruit per unit volume of tree canopy) improved only in the RWB treatment over the control. Total soluble solids and the fruit color index were enhanced by root restriction.

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Ockert P.J. Stander, Jade North, Jan M. Van Niekerk, Tertia Van Wyk, Claire Love, and Martin J. Gilbert

This study aimed to determine the effects of different types of nonpermanent netting (NPN) on foliar spray deposition, insect pest prevalence, and production and fruit quality of ‘Nadorcott’ mandarin (Citrus reticulata) trees in a commercial orchard at Citrusdal (lat. 32 32′31″S, long. 19 0′42″E), Western Cape, South Africa. The deposition quantity (FPC%) of foliar spray volumes of 3500, 7000, or 15,000 L·ha−1 was greater for leaves of control trees compared with leaves treated with NPN during summer (January) (8.8 vs. 6.1; P = 0.0055) and winter (June) (4.8 vs. 3.1; P = 0.0035). Deposition uniformity (CV%) was better for control leaves during summer (64.9 vs. 75.2; P = 0.0062) and winter (59.6 vs. 80.5; P = 0.0014), and deposition quality (ICD%) was better during winter (79.4 vs. 84.2; P = 0.0393). There were no differences in FPC%, CV%, and ICD% for fruit when foliar spray volumes of 3500 and 15,000 L·ha−1 were used for the control and NPN treatment groups during winter. However, with a foliar spray volume of 7500 L·ha−1, fruit from the control treatment group had greater FPC% (19.3 vs. 6.1; P = 0.0262), CV% (70.3 vs. 50.9; P = 0.0484), and ICD% (57.1 vs. 79.9; P = 0.0157). There were no differences in macronutrient concentrations between the leaves of trees subjected to control and NPN treatments, but leaf zinc (<81%; P = 0.0317) and iron (<78%; P = 0.0041) concentrations were lower with the NPN treatment. During short NPN treatments, fruit yield was reduced by ≈37% compared with that after control treatment, and longer NPN treatments had no effect on fruit yield. The reduction in fruit yield with NPN was not related to the effects of NPN on foliar spray deposition or to lower leaf micronutrient concentrations. The lower fruit yield during short NPN treatments was most likely caused by fruit drop that was exacerbated by the removal of the NPN. In the long NPN treatment group, fruit damage caused by sunburn was reduced by 17%, but the outer canopy fruit experienced increased wind damage or scarring. Except for the lower titratable acidity content with the shortest NPN treatment and the higher Brix°:TA ratio with two NPN treatments, NPN did not impact other fruit quality attributes. The use of NPN excluded male wild false codling moths (Thaumatotibia leucotreta) (FCM) males; however, it was still possible to capture a very small amount of mass-released sterile FCM and wild fruit flies under the NPN.

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Andrew J. Krajewski and Etienne Rabe

To investigate the effects of bud age on sprouting and flowering, bearing Clementine mandarin trees were hand-pruned at monthly intervals from late spring to fall. This pruning resulted in regrowth bearing axillary buds ranging in age from 9 to 5 months. After winter rest and during the return bloom, sprouting and flowering were assessed on axils on terminally positioned stems of these ages. The proportion of axillary buds sprouting and the number of spring shoots produced by each sprouting axillary site decreased with decreasing bud age. The proportion of axils sprouting one or more inflorescences, and the average number of flowers per stem also decreased with decreasing bud age. The number of axillary sites per stem, also significantly affected sprouting and flowering. Our results demonstrate the potential of hand-pruning to manipulate sprouting and return bloom depending on when in the summer or autumn the trees are pruned.

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Ute Albrecht and Kim D. Bowman

Huanglongbing (HLB) is a destructive disease of citrus in most citrus-producing countries worldwide. The disease, presumably caused by phloem-limited bacteria of the genus Candidatus Liberibacter, affects all known citrus species and citrus relatives with little known resistance. Typical disease symptoms are the production of abnormal-looking fruit and chlorosis or blotchy mottle of the leaves followed at advanced stages by tree decline and death. Trifoliate orange (P. trifoliata L. Raf.) and some of its hybrids reportedly lack distinct disease symptoms despite infection with the pathogen. US-897 is a hybrid of trifoliate orange and ‘Cleopatra’ mandarin (C. reticulata Blanco), the latter being highly susceptible to HLB. This study investigated whether field-grown, naturally infected trees and greenhouse-grown, graft-inoculated seedlings of this genotype display tolerance or resistance to HLB. It was shown that naturally infected US-897 trees exhibited no distinct disease symptoms commonly associated with HLB, except for the occurrence of few mottled leaves in a small percentage of trees. Analysis of fruit and seed from infected trees did not detect any growth reduction or otherwise negative impact on development. Graft-inoculated US-897 seedlings became polymerase chain reaction (PCR)-positive for the pathogen but exhibited a superior performance compared with ‘Cleopatra’ mandarin seedlings, which displayed severe disease symptoms soon after inoculation. Despite infection, most US-897 seedlings did not develop leaf symptoms typical for HLB. Foliar symptoms observed in a small number of plants at later stages of the disease were faint and difficult to discern. Contrary to ‘Cleopatra’ seedlings, growth in stem diameter was only moderately reduced or unaffected in infected US-897 seedlings. The superior performance of US-897 plants in greenhouse and field locations suggest tolerance of this genotype to Ca. L. asiaticus.

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Daniel Valero, Domingo Martínez, Fernando Riquelme, and María Serrano

The polyamines putrescine, spermidine, and spermine (free form) were measured during 48 hours at 20 °C after mechanical damage of `Fortune' and `Clementine' mandarins (Citrus reticulata Blanco.) with forces of 0, 10, 20, and 30 N. In control fruits (without damage), the three polyamine levels were higher in `Fortune' than in `Clementine', and putrescine and spermine levels tended to diminish during the experiment, while spermidine levels increased slightly in `Fortune' and showed a prominent peak in `Clementine' 6 hours after storage. In general, polyamine levels increased in the peel following bruising. The magnitude of this effect differed for both cultivars and applied forces. Putrescine levels were positively correlated with the applied force in `Fortune', while in `Clementine' only the 20 N force was effective. Spermidine levels also increased in `Fortune' following injury, but there was no difference in effects among the three forces used. Only the 10 N force increased spermidine levels significantly in `Clementine'. Spermine levels were slightly higher than control levels after 48 hours, but not higher than initial levels.

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Neusa M.C. Stenzel, Carmen S.V.J. Neves, José C. Gomes, and Cristiane C. Medina

This study reports the performance (yield, tree size, and fruit quality) of 'Ponkan' mandarin (Citrus reticulata Blanco) on seven rootstocks, evaluated for 11 years under Southern Brazil conditions. Trees on C13 citrange had higher cumulative yield for seven harvests than those on trifoliate orange. Cleopatra mandarin, rough lemon, Rangpur lime, Sunki mandarin, and Volkamer lemon rootstocks maintained their values at an intermediate position and did not present any significant difference regarding C13 citrange, and trifoliate orange. Trees on C13 citrange and on trifoliate orange exhibited the lowest alternate bearing index. Cleopatra mandarin induced the greatest canopy volume, but it was not significantly different from Sunki mandarin and rough lemon. The smallest trees were those on Volkamer lemon and trifoliate orange. The highest yield efficiency came from trees on C13 citrange and the smallest on Cleopatra mandarin. Rootstocks did not significantly affect fruit weight.

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Ed Stover, Scott Ciliento, Monty Myers, Brian Boman, John Jackson Jr., and Max Still

Six trials were conducted to determine whether lower spray volumes or inclusion of different surfactants would permit adequate thinning of mandarin hybrids (Citrus reticulata hybrids) at a much lower cost per hectare. Sprays were applied using a commercial airblast orchard sprayer during physiological drop when fruitlets averaged 8 to 16 mm in diameter. Surfactant was always included at 0.05% v/v. NAA always reduced fruit per tree, increased fruit size, and decreased production of smallest size fruit. However, in only three experiments, contrast of all NAA treatments vs. controls indicated increased production of the largest (80–100 fruit per carton) and most valuable fruit. In four of five experiments, comparison of spray volumes of 600 (only examined in three of four experiments), 1200, or 2300 L·ha–1 demonstrated significant fruit size enhancement from all NAA applications. Most individual NAA treatments resulted in fewer fruit per tree, but there were no statistically significant differences between NAA treatments at different spray volumes. In only one of the four experiments, there was a marked linear relationship between spray volume and fruit per tree, yield, mean fruit size, and production of largest fruit sizes. The effects of surfactants (Activator, a nonionic, Silwet L-77, and LI-700) on NAA thinning were tested in both `Murcott' and `Sunburst'. In comparisons between Silwet L-77 and Activator surfactant, one experiment with `Murcott' showed greater fruit per tree and yield reduction from using Silwet, but with a smaller increase in production of largest fruit sizes, whereas in another `Murcott' experiment, Silwet L-77 reduced numbers of smaller fruit size with no increase in production of larger fruit. Based on these findings, current recommendations for NAA thinning of Fla. mandarins are use of spray volume of ≈1100–1400 L·ha–1 on mature trees with proportionally lower volume on smaller trees. These data appear to support use of a nonionic surfactant rather than other tested surfactants in NAA thinning of Florida mandarins. Because experience with NAA thinning of Florida citrus is limited, it is only recommended where the disadvantages of overcropping are perceived to substantially outweigh the potential losses from overthinning.

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Levava Roiz, Uzi Ozeri, Raphael Goren, and Oded Shoseyov

Aspergillus niger B-1 (CMI CC 324626) extracellular RNase (RNase B1) was purified to homogeneity. It was found to contain two isoforms of 32- and 40-kDa glycoproteins, sharing a 29-kDa protein moiety. Optimal RNase activity was observed at 60 °C and pH 3.5. In `Almog' peach [Prunus persica (L.) Batsch (Peach Group) `Almog'] and `Murcott' tangerine (Citrus reticulata Blanco `Murcott') the enzyme inhibited pollen germination and pollen tube growth in vitro as well as in vivo. In field experiments, spray application of the RNase caused a reduction in `Fantasia' nectarine [Prunus persica (L.) Batsch (Nectarine Group) `Fantasia'] fruit set and interfered with embryo development. The biological effect of the RNase may be of horticultural value, due to its potential to control fertilization.