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Amelia Camprubí and Cinta Calvet

The selection of the most effective arbuscular mycorrhizal (AM) fungi for growth enhancement of citrus cultivars used as rootstocks was the first step toward development of an AM inoculation system in citrus nurseries in Spain. AM fungi were isolated from citrus nurseries and orchards in the major citrus-growing areas of eastern Spain. The most common AM fungi found in citrus soils belonged to Glomus species, and G. mosseae (Nicol. & Gerd.) Gerdemann & Trappe and G. intraradices Schenck & Smith were the AM fungi most frequently associated with citrus roots. The most effective fungus for growth enhancement of citrus rootstocks was G. intraradices. Significant differences in mycorrhizal dependency among rootstocks were confirmed. Sour orange (Citrus aurantium L.) and Cleopatra mandarin (C. reshni L.) were more dependent than Troyer citrange [C. sinensis (L.) Obs. × Poncirus trifoliata (L.) Raf.] and Swingle citrumelo (C. paradisi Macf. × P. trifoliata). Moreover, several inoculation systems for plant production were evaluated for their effectiveness in promoting root colonization of the rootstock cultivars.

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Eric H.C. Chilembwe, William S. Castle, and Daniel J. Cantliffe

Commercially processed citrus seeds of Carrizo citrange [Citrus sinensis (L.) Osb. × Poncirus trifoliata (L) Raf.], Swingle citrumelo (C. paradisi Macf. × P. trifoliata), Cleopatra mandarin (C. reticulate Blanco), and sour orange (C. aurantium L.) were used to test the effects of grading, hydrating, and priming on the rate of germination and seedling emergence. Sorting seed into groups by fresh weight or diameter did not generally improve seed performance. Seed fresh weight was highly correlated with maximum seed diameter; also, large seed weight and size were associated with a larger number of embryos. When seedlings from the extra embryos were removed, large seed produced the largest seedlings. Soaking seeds in aerated water significantly increased germination and emergence rates over unsoaked seeds. Soaking at 35C rather than 25C enhanced these differences. Priming seeds in one of three solutions of polyethylene glycol 6000 (—0.6 to—1.2 Mpa) was not successful> as germination and emergence Per centages were lower than in distilled water.

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Charles A. Powell, Phyllis A. Rundell, and Robert R. Pelosi

Bark chips from six container-grown citrus trees, infected with nondecline-inducing citrus tristeza virus (CTV) isolates and maintained in a vector-free greenhouse for 10 years, 15 commercial grapefruit (Citrus paradisi Macf.) trees, and 16 commercial sweet orange [C. sinensis (L.) Osbeck] trees were used to inoculate three indicator plants each of `Madam Vinous' sweet orange [C. sinensis (L.) Osbeck], sour orange (C. aurantium L.), `Duncan' grapefruit (C. paradisi Macf.), `Mexican' lime [C. aurantifolia (Christm.)], Swingle citrumelo [C. paradisi Macf. × Poncirus trifoliota (L.) Raf.], and sour orange grafted with `Hamlin' sweet orange [C. sinensis (L.) Osbeck]. All plants providing bark chips had repeatedly tested positive by enzyme-linked immunosorbent assay (ELISA) for CTV [reacted with monoclonal antibody (MAb) 17G11], but tested negative for Florida decline-inducing isolates of CTV (did not react with MAb MCA13). After 6 months in vector-free greenhouses, all in oculated trees (except Swingle citrumelo, which is considered CTV resistant) were positive for CTV by 17G11 ELISA. In addition, some indicator plants inoculated from nine (two container, two commercial grapefruit, and five commercial orange trees) of the 37 bark chip source trees also were positive for decline-inducing CTV by MCA13 ELISA. Some of these positive indicators also showed vein-clearing symptoms characteristic of infection with a severe isolate of CTV. No control, noninoculated indicators in the same greenhouse, became infected with either decline-inducing or nondecline-inducing CTV. These results indicate that decline-inducing isolates of CTV can be present as a minor component of a mixture at levels undetectable by ELISA, and that these decline-inducing isolates can become detectable by ELISA and sometimes by symptoms when inoculated into indicator plants.

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O. Gulsen and M.L. Roose

Chloroplast DNA (cpDNA) restriction fragment length polymorphisms were used to study the parentage of lemons [Citrus limon (L.) Burm. f.] and several other putative hybrids. The 30 citrus accessions studied included nine lemons, three pummelos [C. maxima (Burm.) Merrill], three citrons (C. medica L.), three mandarins (C. reticulata Blanco, C. clementina Hort. Ex. Y. Tan., and C. sunki Hort. ex. Tan.), and single representatives of 13 other taxa. Four different fragments of citrus cpDNA were amplified by polymerase chain reaction using four universal chloroplast primers and amplification products were digested with four endonucleases, Rsa I, Dra I, Hae III, and Mbo I. A total of 104 different restriction fragments were scored and used for phylogenetic analysis by parsimony. The three taxa which have been proposed as ancestral Citrus L. sp., C. medica, C. maxima, and C. reticulata, had distinct cpDNA patterns. Pummelo contributed the chloroplast genome to lemons, sweet orange [C. sinensis (L.) Osbeck], and Bergamot orange (C. bergamia Risso and Poit.), and mandarin contributed the chloroplast genome to rough lemons (C. jambhiri Lush.), Rangpur lime (C. limonia Osbeck), and `Mexican' lime [C. aurantifolia (Christm.) Swing.]. Data suggest that the particular accessions of these ancestral species that were studied were not directly involved in the hybridization events that created these hybrids, since they have similar but not identical cpDNA restriction fragments. Using inter-simple sequence repeat markers that amplified from nuclear genomic DNA, a set of samples including pummelos, citrons, mandarins, sour oranges (C. aurantium L.), and `Samuyao' papeda (C. micrantha Wester) were tested as possible parents of lemons. Sour orange and citron together had all nuclear and chloroplast fragments found in lemon and are therefore proposed to be the maternal and paternal parents, respectively, of many commercial lemon cultivars, including `Lisbon', `Eureka', `Villafranca', and `Monachello'.

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G.R. Stino, A.E. Abou Aziz, A.A. Elezaby, and E.A. Abd Elmoneim

Valencia orange trees [Citrus sinensis (L.) Osb.], budded on sour orange (C. aurantium) rootstock, were sprayed with four different potassium salts during 1995 and 1996. Twenty 5-year-old trees were sprayed once per month with one of the following material. K0: distilled water, K1: potassium green (a compound of different potassium salts, 35% K2O), K2: potassium nitrate (35% K2O), K3: potassium citrate (35% K2O), or K4: potassium sulfate (51% K2O). The vegetative growth occurred in three distinct successive cycles, i.e., spring, summer, and autumn. These cycles differed in time of commencement, duration and termination with respect to season. All potassium treatments significantly increased the shoot length and number of leaves/shoot for the three growth cycles. However, potassium green was superior overall other treatments. Percentage of leaf miner infestation was reduced at all potassium treatments. Potassium applications significantly increased leaf contents of N, P, K and decreased Ca and Mg levels.

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Mongi Zekri

Salt tolerance of Carrizo citrange (CC), sour orange (SO), and Cleopatra mandarin (CM) rootstocks during seedling emergence (SE) and early seedling growth (SG) was studied under greenhouse conditions. Increasing salt (NaCl + CaSO4) concentration delayed and emergence, reduced SG, but did not affect emergence spread. At the first salinity level (5 mmol), emergence of the first seedling (EFS) was delayed by 2 to 3 days in CC and one day in SO but was not affected in CM. At the highest salt level (80 mmol), EFS was delayed by up to 6, 7, and 5 days for CC, SO, and CM, respectively. At the two lowest salinity levels (5 and 10 mmol), final percent emergence (FPE) was not affected in CC but was reduced in SO and CM while shoot biomass was reduced in CC but was not affected in SO and CM. At the 80 mmol salt level, FPE was reduced by 23% in CC and by 33% in SO and CM while SG was reduced by 70% in CC and by 60% in SO and CM. Among the rootstocks studied, the delay in emergence was not necessarily more salt sensitive than FPE. However, SG was generally more affected by salinity than SE, particularly at high salinity levels (20, 40, and 80 mmol).

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Leah E. Willis, Frederick S. Davies, and D.A. Graetz

One-year-old `Hamlin' orange [Citrus sinensis (L.) Osb.] trees on sour orange rootstock (C. aurantium L.) were used to compare various fertigation frequencies and rates with application of granular materials. In Expt. 1, granular fertilizer was applied five times per year or liquid fertilizer was applied five, 10, or 30 times per year at 0.23 kg N/tree per year as an 8N-3.4P-6.6K formulation. In Expt. 2, an additional treatment of granular and liquid material was applied three times per year, but fertilizer rate and formulation were the same as in Expt. 1. Experiment 3 included the same application frequencies as Expt. 1, but with two rates of N (0.11 or 0.06 kg N/tree per year). Soil samples were taken from each treatment 1, 4, and 7 days after fertilization at depths of 0-15, 16-46, and 47-76 cm for nutrient analyses. Trunk diameter, shoot growth, and tree height were similar for all treatments 8 months after planting in Expts. 1 and 2, while trees in Expt. 3 had significantly less growth at the lower rate. Soil NH4-N and NO3-N concentrations for all liquid treatments within 1 week of fertilization were highest for the five times per year treatment at the 0- to 15-cm depth, but nutrient concentrations of all liquid treatments were similar at the other depths. For most dates and depths, NH4-N and NO3-N concentrations were similar for both fertilizer rates.

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C. Gregoriou and C.V. Economides

Growth, yield, and fruit quality were recorded for Ortanique tangor (Citrus reticulata Blanco) on 11 rootstocks until the trees were 12 years old. Trees on Volkameriana (C. volkameriana Pasq.), rough lemon, and `Estes rough lemon (C. jambhiri Lush.) were more productive per unit of tree size, and their cumulative yields per tree were significantly higher than those of trees on the other rootstocks. There was no significant difference between cumulative yields of Ortanique on the following rootstocks: sour orange (C. aurantium L.), `Palestine' sweet lime (C. limettioides Tan.), `Red' rough lemon (C. jambhiri Lush.), Rangpur (C. limonia Osbeck), and Amblycarpa (C. limonellus var. amblycarpa Hassk.). However, yield on these rootstocks was significantly higher than on Carrizo and Troyer citranges [C. sinensis (L.) Osbeck × Poncirus trifoliata (L.) Raf.] and `Swingle' citrumelo [C. paradisi Macf. × P. trifoliata (L.) Raf.]. The high productivity per unit of tree size of `Palestine' sweet lime suggested that this rootstock could be used advantageously in closely spaced plantings. Rootstocks affected fruit size, weight, rind thickness, juice content, total soluble solids concentration (SSC), and total acids, but the differences were not large enough to be of practical importance.

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J.W. Grosser, J. Jiang, E.S. Louzada, J.L. Chandler, and F.G. Gmitter Jr.

Production of tetraploid somatic hybrids that combine complementary diploid rootstock germplasm via protoplast fusion has become a practical strategy for citrus rootstock improvement, with the overall objective of packaging necessary disease and pest resistance into horticulturally desirable, widely adapted rootstocks. Citrus somatic hybridization techniques have been advanced to the point where numerous somatic hybrid rootstocks can now be produced and propagated for evaluation on a timely basis. Herein we report the production of 11 new somatic hybrid rootstock candidates from 12 different parents, including Milam lemon hybrid (Citrus jambhiri Lush.), Cleopatra mandarin (C. reticulata Blanco), sour orange (C. aurantium L.), `Succari' sweet orange [C. sinensis (L.) Osbeck], `Redblush' grapefruit (C. paradisi Macf.), `Nova' tangelo [C. reticulata × (C. paradisi × C. reticulata)], `Kinkoji' (C. obovoidea Hort. Ex Takahashi), Swingle citrumelo [C. paradisi × Poncirus trifoliata (L.) Raf.], Carrizo citrange (C. sinensis × P. trifoliata), rough lemon 8166 (C. jambhiri), and Palestine sweet lime (C. limettoides Tan.). All hybrids were confirmed by cytological and VNTR-PCR analyses, and have been propagated, budded with a commercial scion, and field-planted for performance evaluation.

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Sudahono, D.H. Byrne, and R.E. Rouse

Eighteen citrus rootstock seedling lines were tested for their tolerance to Fe chlorosis using sand culture. Potassium carbonate was used to induce Fe-deficiency chlorosis. Chlorosis was quantified by 1) visual ratings, 2) SPAD-502 chlorophyll meter readings, 3) leaf chlorophyll concentration, 4) leaf active Fe, and 5) leaf total Fe. The first four criteria were well correlated among each other but not with leaf total Fe. Although any of the first four measurements could be used to quantify chlorosis, visual ratings and SPAD-502 readings were more convenient. The rootstock that have been reported to be tolerant or very susceptible to Fe chlorosis in calcareous soils were rated similarly for tolerance to bicarbonate-induced Fe chlorosis. Nontrifoliate types such as Texas sour orange (C. aurantium L.), Cleopatra mandarin (C. reticulata Blanco), Vangasay lemon (C. limon Burro.), and Ridge pineapple x Milam 1578-201 (C. sinensis L. Osbeck x C. jambhiri) were tolerant to moderately tolerant. Although most of the trifoliate hybrids tested were moderately susceptible to very susceptible, Smooth Seville x Argentine trifoliate {[C. grands (L.) Osbeck x C. aurantium] x Poncirus trifoliata (L.) Raf.} and F-81-12 citrange (C. sinensis x P. trifoliata) exhibited relatively high tolerance to lime-induced Fe chlorosis.