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Sandra B. Wilson, Robert L. Geneve, and Fred T. Davies

Interactive web-based questions were developed for students to review subject matter learned in an online plant propagation course. Articulate Storyline software was used to build nearly 250 review questions with five different testing styles to ascertain proficiency in subject areas, including the biology of propagation, the propagation environment, seed propagation, vegetative propagation, micropropagation, and cell culture. Questions were arranged to correspond to the supporting textbook chapters in Hartmann and Kester’s Plant propagation: Principles and practices, ninth edition. These are open access and available to instructors and students worldwide. Users received immediate feedback for each question answered correctly or incorrectly. The system remembers where one leaves off, which enables starting and stopping multiple times within a chapter. Means of pre- and posttest responses to nine content knowledge items showed that students perceived a significant content knowledge gain in the course. These online interactive reviews can be adapted easily to other courses in a variety of fields, including horticulture, botany, systematics, and biology. They can also be expanded to overlay multiple objects and trigger events based on user response. Since inception, the website hosting these online reviews averaged 156 unique visitors per month. Students have reported this to be a useful tool to prepare them for course exams.

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Yifei Wang, Stephanie K. Fong, Ajay P. Singh, Nicholi Vorsa, and Jennifer Johnson-Cicalese

The flavonoid and organic acid profiles of one cultivated tetraploid and six wild diploid blueberry species (Vaccinium spp.) were systematically investigated using high-performance liquid chromatography–electrospray ionization–tandem mass spectrometry (HPLC-ESI-MS-MS). Eighteen individual anthocyanins from five aglycone classes were characterized among species, with malvidin and delphinidin glycosides accounting for 31.4% and 29.1% of total anthocyanins. Twenty-three flavonol glycosides from six aglycone classes were identified, among which quercetin and myricetin glycosides accounted for more than 80% of total flavonols in most species. Both inter- and intraspecies differences in anthocyanin and flavonol composition were observed, as described by principal component analysis. Only B-type proanthocyanidins were found in blueberry species, and highly polymerized molecules with degree of polymerization greater than 10 appeared to be the most abundant fraction. Although overall proanthocyanidin levels varied from 27.7 to 146.3 mg/100 g fruit, all species exhibited similar proanthocyanidin composition. Citric, quinic, and shikimic acid were the major identified blueberry organic acids. However, their relative abundance varied across species. In certain species either citric acid (e.g., Vaccinium darrowii) or quinic acid (e.g., Vaccinium corymbosum) was lacking.

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Samia Lotfy, Francois Luro, Françoise Carreel, Yann Froelicher, Delphine Rist, and Patrick Ollitrault

Somatic hybridization allows the creation of new patterns of nuclear, mitochondrial and chloroplastic association. It is therefore necessary to master cytoplasmic molecular markers to determine the genetic origin of both organelles of plantlets obtained from protoplasts fusion. In the case of Citrus and related genera, only southern blot hybridization and restriction fragment-length polymorphism (RFLP) techniques were used for this task until now. Here, we describe the use in the Aurantioideae subfamily, of a simple and non labeling cleaved amplified polymorphic sequence (CAPS) technique, to determine the cytoplasmic genome origin of intergeneric somatic hybrids. Mitochondrial and chloroplastic universal primers previously selected for population genetic studies in Quercus by Demesure et al. (1995) are used with some modifications. The variability of cytoplasmic genome among somatic fusion partners is detected by coupling amplification and restriction reactions. Digested DNA fragments are analyzed by agarose gel electrophoresis (PCR-RFLP). This technique has been applied for the analysis of the cytoplasmic constitution of somatic hybrids arising from intergeneric, intersubtribal and intertribal combinations. Systematic transmission of the mitochondria from protoplasts isolated from embryogenic callus parents was confirmed.

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Kelly T. Morgan, T.A. Obreza, and J.M.S. Scholberg

Understanding the growth pattern of fibrous, orange tree [Citrus sinensis (L.) Osbeck] roots enables proper fertilizer placement to improve nutrient uptake efficiency and to reduce nutrient leaching below the root zone. The objective of this study was to develop relationships defining citrus fibrous root length density (FRLD) as a function of soil depth, distance from the tree trunk, and tree size. Root systems of 18 trees with tree canopy volumes (TCV) ranging from 2.4 to 34.3 m3 on two different rootstocks and growing in well-drained sandy soils were sampled in a systematic pattern extending 2 m away from the trunk and 0.9 m deep. Trees grown on Swingle citrumelo [Citrus paradisi Macf. × Poncirus trjfoliata (L.) Raf.] rootstock had significantly greater FRLD in the top 0.15 m than trees on Carrizo citrange (C. sinensis × P. trifoliata). Conversely, Carrizo citrange had greater FRLD from 0.15 to 0.75 m below the soil surface. FRLD was significantly greater for ‘Hamlin’ orange trees grown on Swingle citrumelo rootstock at distances less than 0.75 m from the tree trunk compared with those on Carrizo citrange. Fibrous roots of young citrus trees developed a dense root mat above soil depths of 0.3 m that expanded both radially and with depth with time as trees grow and TCV increased. Functional relationships developed in this study accounted for changes in FRLD with increase in tree size.

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Janel L. Giovannelli, Mark W. Farnham, Min Wang, and Allan E. Strand

Downy mildew, caused by the fungal parasite Peronospora parasitica (Pers.: Fr.) Fr., is a destructive disease of Brassica oleracea L. crops, including broccoli (B. oleracea, Italica Group). The development and deployment of downy mildew resistant broccoli cultivars is a priority for breeders and producers. Identification of genetic markers linked to downy mildew resistance genes should facilitate selection for resistance and pyramiding of resistance genes into cultivars. The objectives of this study were to 1) identify RAPD markers linked to a single dominant gene for resistance in broccoli, 2) clone and sequence the linked RAPD markers, and 3) develop and evaluate SCAR markers as screening tools for resistance. Bulked segregant analysis led to the identification of eight linked RAPD markers following a screen of 848 decamers. Two of the linked RAPD fragments, UBC359620 and OPM16750, were converted to dominant SCAR markers linked in coupling to the resistance locus at 6.7 and 3.3 cM, respectively. The SCAR marker based on UBC359620 sequence exhibited less accuracy (94%) than the original RAPD (96%) in differentiating resistant and susceptible plants, but the accuracy (97%) of the OPM16750-SCAR was not different than the original RAPD. These SCAR markers are among the first genetic markers found linked to a gene conferring cotyledon-stage downy mildew resistance in B. oleracea. Results of this work provide breeders with useful information and tools for the systematic development of resistant cultivars.

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A. Medlicott, J. Brice, T. Salgadol, and D. Ramirez

No systematic curing and storage techniques are currently used with onions in Honduras; postharvest losses occur rapidly. The objective of this study was to evaluate the use of storage bins (maximum capacity 7t) that use forced ambient air ventilation to manipulate the atmospheric conditions around the onions. The desired storage conditions were 26 to 30C and 60% to 75% relative humidity. Ventilation regimes were manipulated in an attempt to obtain these conditions. The rate of deterioration in four varieties of onions over a 3-month period was determined and compared with onions stored under normal ambient conditions. Marketable onions in the forced-air storage bin compared to the controls stored under ambient conditions after 13 weeks were 82% vs. 37% for `Granex 33'; 71% vs. 40% for `Granex 429'; 63% vs. 31% for `Granex 438'; and 90% vs. 44% for `Texas Grano 502'. This represents a significant increase in the number of marketable onions after storage. All losses were increased by rain and tornado damage after 1 month of storage. The methods used to maintain uniform temperature and humidity conditions in the storage bin are discussed together with the problems encountered. The construction and operating costs are given together with the market prices and the required returns to cover the bin costs.

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Mark W. Farnham

A collection of collard (Brassica oleracea L., Acephala group) germplasm, including 13 cultivars or breeding lines and 5 landraces, was evaluated using randomly amplified polymorphic DNA (RAPD) markers and compared to representatives of kale (Acephala group), cabbage (Capitata group), broccoli (Italica group), Brussels sprouts (Gemmifera group), and cauliflower (Botrytis group). Objectives were to assess genetic variation and relationships among collard and other crop entries, evaluate intrapopulation variation of open-pollinated (OP) collard lines, and determine the potential of collard landraces to provide new B. oleracea genes. Two hundred nine RAPD bands were scored from 18 oligonucleotide decamer primers when collard and other B. oleracea entries were compared. Of these, 147 (70%) were polymorphic and 29 were specific to collard. Similarity indices between collard entries were computed from RAPD data and these ranged from 0.75 to 0.99 with an average of 0.83. Collard entries were most closely related to cabbage (similarity index = 0.83) and Brussels sprouts entries (index = 0.80). Analysis of individuals of an OP cultivar and landrace indicated that intrapopulation genetic variance accounts for as much variation as that observed between populations. RAPD analysis identified collard landraces as unique genotypes and showed them to be sources of unique DNA markers. The systematic collection of collard landraces should enhance diversity of the B. oleracea germplasm pool and provide genes for future crop improvement.

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Marissa Moses and Pathmanathan Umaharan

Capsicum chinense is commercially the most important pepper species grown in the Caribbean. It is popularly used to impart pungency and flavor to Caribbean cuisine. However, unlike Capsicum annuum, which is the most commercially exploited domesticated species internationally, C. chinense has not been methodically collected or characterized for systematic improvement through plant breeding. The objectives of the study were to assess the diversity of C. chinense and its structure within the Caribbean basin and to determine its phylogenetic relationship to groups within South America. DNA isolated from 201 accessions of C. chinense, representing geographical regions where the species is found, were amplified using arbitrary primers to generate 138 polymorphic and reproducible random amplification of polymorphic DNA (RAPD) markers. Nei’s and Shannon’s diversity indices for C. chinense (0.28 and 0.419, respectively) were higher in South America compared with Central America or the Caribbean, corresponding to its putative center of diversity. The study showed the existence of three phylogenetic clusters within C. chinense. The largest cluster consisted of accessions from the Upper Amazon region, the Guianas including Venezuela, and the Lesser Antilles of the Caribbean. The other major cluster was represented by accessions principally from the Lower Amazon region. Another distinct but small cluster consisted of samples solely from the Greater Antilles of the Caribbean. The discovery of the three phylogenetic clusters within C. chinense may have potential for exploiting heterosis in breeding. The implications of the findings to the understanding of the phylogenetic origin and distribution of C. chinense are discussed.

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Christopher M. McGuire

1 Current address: Section of Ecology and Systematics, Cornell Univ., Ithaca, NY 14853. I thank Ian Merwin, John Ray, Marvin Pritts, and Mary Jo Kelly for advice and assistance, and my family for encouragement and

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Rebecca L. Loughner, Daniel F. Warnock, and Raymond A. Cloyd

Foundation. The authors would like to thank Stephanie Larsen and Andreana Lau for their assistance with this research and David A. Nickle, USDA Systematic Entomology Laboratory, Beltsville, Md., for the identification of thrips species used in this project