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Peng Ling, Fred G. Gmitter Jr., Larry W. Duncan, and S. Y. Xiao

A family of 63 citrus intergenemic backcross hybrids was used for this study. The parents and hybrids were multiplied by rooted cuttings, with 6 uniform replicates selected per hybrid, and each plant was inoculated with citrus nematodes (Tylenchulus semipenetrans) 5 times over 2 mo. The number of nematode female larvae per gram of fine fresh root was determind 2 mo after the last inoculation. The phenotypic variation of the hybrids was continuous and wide-ranged, from 8.0 females· g-1 of root tissue (resistant parent Swingle citrumelo=15.6) to 620.0 females· g-1 of root tissue (susceptible parent LB 6-2=540.5). Bulked segregant analysis (BSA), using RAPD fragments, was conducted with 2 DNA bulks of individuals from the extremes of the phenotypic distribution. Three hundred twenty primers were screened and 5 were found to generate repeatedly single RAPD fragments specific to the resistant bulk. The segregation of resistance-associated fragments among the individuals was examined, and the linkage between these markers and potential nematode resistance loci was estimated.

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Xing-Zheng Fu, Fei Xing, Li Cao, Chang-Pin Chun, Li-Li Ling, Cai-Lun Jiang, and Liang-Zhi Peng

To compare the effects of various zinc (Zn) foliar fertilizers on correcting citrus Zn deficiency and to explore an effective correcting method, three common Zn fertilizers, Zn sulfate heptahydrate (ZnSO4.7H2O), Zn chloride (ZnCl2), and Zn nitrate hexahydrate [Zn(NO3)2.6H2O], were selected to spray the Zn-deficient citrus leaves, tested at different concentrations, with or without organosilicone surfactant. Zn content, chlorophyll levels, and photosynthesis characteristics of leaves were analyzed. Leaf Zn content was significantly increased with increase of the sprayed Zn concentration of the three Zn fertilizers. However, when the sprayed Zn concentration of ZnSO4.7H2O exceeded 200 mg·L−1, and Zn concentration of ZnCl2 or Zn(NO3)2.6H2O exceeded 100 mg·L−1, obvious necrotic spots formed on leaves. This necrosis disappeared when 0.025% organosilicone was added to the three Zn fertilizer solutions, even at a Zn concentration of 250 mg·L−1. Meanwhile, the Zn contents of leaves increased one to four times for these treatments. Furthermore, foliar application of the three Zn fertilizers significantly improved chlorophyll levels and photosynthetic capacity of Zn-deficient leaves. The data of chlorophyll and photosynthesis characteristics indicate that the correcting effect of ZnCl2 and Zn(NO3)2.6H2O is better than that of ZnSO4.7H2O, and could be further improved via supplement of organosilicone. In conclusion, ZnCl2 or Zn(NO3)2.6H2O containing 250 mg·L−1 of Zn and supplemented with 0.025% organosilicone is a safe and effective formulation of Zn foliar fertilizer for correcting citrus Zn deficiency.

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Aditi D. Satpute, Chunxian Chen, Fredrick G. Gmitter Jr., Peng Ling, Qibin Yu, Melinda R. Grosser, Jude W. Grosser, and Christine D. Chase

In cybridization, new combinations of nuclear and cytoplasmic genes result in a unique genotype that may bring cellular, physical, physiological, and biochemical changes to the plant. This has been demonstrated in the unexpected cybrids generated from the fusion of citrus (Citrus sp.) protoplasts in two independent experiments. The first experiment was conducted to generate potentially seedless triploids by fusing diploid protoplasts of embryogenic ‘Dancy’ mandarin (Citrus reticulata) suspension culture cells with haploid ‘Ruby Red’ grapefruit (C. paradisi) protoplasts derived from tetrad-stage microspores. After multiple attempts, only one triploid was recovered, but several diploid plants with typical grapefruit morphology were also regenerated. In the second experiment, protoplasts derived from embryogenic ‘Dancy’ mandarin suspension culture were fused with nonembryogenic protoplasts from ‘Duncan’ grapefruit leaves in an effort to produce an allotetraploid somatic hybrid. The fruit from the resulting trees resembled grapefruit in morphology and type, and maintained excellent quality throughout the summer, when commercial grapefruit rapidly loses quality. Fruit on these trees remained firm with exceptional sweetness and good flavor into August, and without seed germination. The regenerants obtained in the protoplast fusion experiments were confirmed as cybrids by genetic marker analyses. The test grapefruit were identical to commercial ‘Ruby Red’ grapefruit at six nuclear simple sequence repeat (SSR) marker loci, but identical to ‘Dancy’ with respect to a mitochondrial intron marker. The plastid genomes of individual trees originated from either fusion partner. In the first experiment, haploid ‘Ruby Red’ protoplast preparations must have also contained contaminant diploid protoplasts. Apart from the value of altered fruit quality attributes in the marketplace, these plants provide an opportunity to understand the contributions of cytoplasmic organelle genetics to important citrus fruit-breeding objectives.