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Alvin M. Simmons and Amnon Levi

The B-biotype sweetpotato whitefly, Bemisia tabaci (Gennadius), feeds on and damages numerous vegetable crops including watermelon (Citrullus sp.). Seven watermelon cultivars, a triploid line, and 16 U.S. Plant Introduction accessions (PIs) of C. lanatus var. lanatus; 10 PIs of C. lanatus var. citroides; and eight PIs of C. colocynthis, were evaluated for resistance to B. tabaci. Bioassays were based on nonpreference and performance of the whiteflies on the 42 Citrullus genotypes. Most of the watermelon cultivars and C. lanatus PIs tested were highly susceptible to whitefly infestation, while the C. colocynthis accessions exhibited whitefly resistance. Among the C. colocynthis accessions tested, PI 386015, PI 386018, and PI 386024 were most resistant to B. tabaci. This study identified useful sources of germplasm that can be used for the improvement of watermelon for resistance to whiteflies.

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Amnon Levi and Claude E. Thomas

A genetic linkage map was previously constructed for watermelon using a wide testcross population [{Plant Accession Griffin 14113; Citrullus lanatus var. citroides (L.H. Baiely) Mansf.} × the watermelon cultivar New Hampshire Midget; NHM {(Citrullus lanatus (Thunb.) Matsum. & Nakai var. lanatus)} × United States Plant Introduction (PI) 386015 {Citrullus colocynthis (L.) Schrad.}]. One-hundred forty-six markers [randomly amplified polymorphic DNA (RAPD), intersimple sequence repeat (ISSR), amplified fragment length polymorphism (AFLP), and sequence-related amplified polymorphism (SRAP) markers] unique to NHM and representing different linkage groups on the map were tested for polymorphism among 24 watermelon cultivars limited in genetic diversity. Five (9.4%) of 53 RAPD, six (40.0%) of 15 ISSR, 30 (81.0%) of 37 AFLP, and 33 (80.5%) of 41 SRAP markers tested produced polymorphism among the 24 cultivars. The polymorphic markers used in this study are scattered throughout the watermelon genome. However, a large number (19 of the 30) of AFLP markers clustered on one linkage group on the map. The SRAP markers proved to be most effective in producing polymorphism and in representing different linkage regions of watermelon genome. The polymorphic markers represent all 10 large linkage groups and five of the nine small linkage groups (altogether 15 of 19 linkage groups) of the genetic linkage map constructed so far for watermelon. These polymorphic markers can be useful in DNA fingerprinting of cultivars, in testing seed purity of breeding lines, and in identifying triploid (seedless) hybrid watermelons derived from crosses between closely related tetraploid and diploid lines.

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Judy A. Thies and Amnon Levi

Root-knot nematodes [Meloidogyne arenaria (Neal) Chitwood, Meloidogyne incognita (Kofoid & White) Chitwood, and Meloidogyne javanica (Treub) Chitwood] are serious pests of watermelon [Citrullus lanatus (Thunb.) Matsum. & Nakai var. lanatus] in the southern United States and worldwide. Watermelon cultivars with resistance to any of these nematode pests are not available. Therefore, we evaluated all accessions of Citrullus colocynthis (L.) Schrad.(21) and Citrullus lanatus (Thunb.) Matsum. & Nakai var. citroides (L.H. Bailey) Mansf.(88), and about 10% of C. lanatus var. lanatus (156) accessions from the U.S. Plant Introduction (PI) Citrullus germplasm collection for resistance to M. arenaria race 1 in greenhouse tests. Only one C. lanatus var. lanatus accession exhibited very low resistance [root gall index (GI) = 4.9] and 155 C. lanatus var. lanatus accessions were susceptible (GI ranged from 5.0 to 9.0, where 1 = no galls and 9 = ≥81% root system covered with galls). All C. colocynthis accessions were highly susceptible (GI range = 8.5 to 9.0). However, 20 of 88 C. lanatus var. citroides accessions were moderately resistant with a GI range of 3.1 to 4.0; overall GI range for the C. lanatus var. citroides accessions was 3.1 to 9.0. Resistance to M. arenaria race 1 identified in the C. lanatus var. citroides accessions was confirmed on a subset of accessions in a replicated greenhouse test. The results of our evaluations demonstrated that there is significant genetic variability within the U.S. PI Citrullus germplasm collection for resistance to M. arenaria race 1 and also identified C. lanatus var. citroides accessions as potential sources of resistance.

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Judy A. Thies and Amnon Levi

Root-knot nematodes (Meloidogyne spp.) cause extensive damage to watermelon [Citrullus lanatus (Thunb.) Matsum. & Nakai var. lanatus], and resistance to root-knot nematodes has not been identified in any watermelon cultivar. Twenty-six U.S. Plant Introductions (PIs) of Citrullus lanatus (Thunb.) Matsum. & Nakai var. citroides (L. H. Bailey) Mansf., one PI of C. lanatus var. lanatus, and three PIs of Citrullus colocynthis (L.) Schrad. were evaluated in greenhouse tests for resistances to Meloidogyne incognita (Kofoid & White) Chitwood race 3 and Meloidogyne arenaria (Neal) Chitwood race 2. Twenty-three of the C. lanatus var. citroides PIs and the C. lanatus var. lanatus PIs were previously identified as moderately resistant to M. arenaria race 1. Overall, the C. lanatus var. citroides PIs exhibited low to moderate resistance, and the C. lanatus var. lanatus and C. colocynthis PIs were susceptible to both M. incognita race 3 and M. arenaria race 2. The C. lanatus var. citroides PI 482303 was the most resistant PI with gall index (GI) = 2.88 and reproductive index (RI) = 0.34 for M. incognita race 3 and GI = 3.46 and RI = 0.38 for M. arenaria race 2 (1 = no galling; 5 = 26% to 38% root system galled; 9 = 81% to 100% root system galled). These results demonstrate that there is significant genetic variability within C. lanatus var. citroides for reaction to M. incognita and M. arenaria race 2, and several C. lanatus var. citroides PIs may provide sources of resistance to root-knot nematodes.

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Kai-Shu Ling and Amnon Levi

One-hundred ninety U.S. PIs of bottlegourd [Lagenaria siceraria (Mol.) Standl.] were evaluated for their resistance to the Florida strain of Zucchini yellow mosaic virus (ZYMV-FL). Seedlings in the first leaf stage were mechanically inoculated with freshly prepared ZYMV-FL tissue extract in a greenhouse. Four weeks postinoculation, plants were visually evaluated for symptom expression and tissue samples from upper noninoculated leaves were collected for serological analysis with enzyme-linked immunosorbent analysis (ELISA). A combination of symptom expression and ELISA value was considered in determining the resistance or susceptibility for each accession. Of the 190 L. siceraria PIs screened, 36 accessions were in complete resistance (no disease symptom with negative ELISA on all tested plants), 64 PIs showed partial resistance (some of the tested plants were resistant, whereas others were susceptible), and 90 PIs were susceptible (severe symptom and positive ELISA on all tested plants). The ZYMV-FL resistance exists mostly among L. siceraria PIs collected in India. Thirty-three of the 36 L. siceraria PIs showing ZYMV-FL resistance were collected in India, one in Indonesia, one in South Africa, and one in Zimbabwe. To rule out any potential escapes in the primary screening, a repeated test using representative accessions, including 10 susceptible, three partially resistant, and three completely resistant PIs, was done to confirm the ZYMV-FL resistance. Furthermore, the resistance to ZYMV-FL was shown to be heritable in progenies generated through self-pollination of single plants in each of five resistant PIs as well as in three F1 hybrids.

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Hazel Y. Wetzstein and Amnon Levi.

Embryogenesis in higher plants follows a standard developmental program with sequential stages of histodifferentiation, maturation (reserve deposition), and postabscission (desiccation and rapid decline in metabolic activity). In this study, morphological, physiological and anatomical characteristics were integrated to demarcate the developmental stages of pecan embryos. Fruit were collected, morphological characteristics were recorded, fresh and dry weights, and water content of embryos were determined, and embryos were prepared for microscopic study. The procedures used here can be a useful guide for characterizing embryo development in pecan and related species.

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Claude E. Thomas, Amnon Levi and Ellis Caniglia

Two hundred sixty-six Citrullus lanatus (Thumb.) Matsum. & Nakai accessions (Plant Introductions and named cultivars) were tested against a race 2 Sphaerotheca fuliginea (Schlechtend.: Fr.) Pollacci isolate to evaluate for resistance to powdery mildew disease. Growth room-grown seedlings were artificially inoculated with conidia from watermelon host leaves at 2-day intervals from the appearance of the first true leaf until test results data were taken, when the second true leaf was fully expanded. Plants were evaluated on a 1 to 9 scale of increasing disease severity. Disease indices (DIs) were calculated as weighted averages for each entry. All genotypes with resistant plants (powdery mildew rating 1 to 3) were reevaluated in a replicated test of 3 replications of 10 plants each. Disease indices were again calculated. Twenty-two plant introductions (PIs) and one named variety displayed intermediate resistance to powdery mildew in the replicated test with DIs ranging from 5.0 to 6.0.

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Amnon Levi, Elizabeth Ogden and Lisa J. Rowland

Efforts are underway to develop genetic linkage maps for two interspecific blueberry populations (Vaccinium darrowi × V. elliottii and V. caesariense-derived populations). To date, 72 RAPD markers have been mapped, and another 200 markers have been identified as suitable for mapping in the V. darrowi × V. elliottii-derived population. Inheritance of 40 RAPD markers has been followed, and additional 40 RAPD markers have been identified as suitable for mapping in the V. darrowi × V. caesariense population. These two populations are comprised of individual plants that should have a wide range of chilling requirements. At a later date, plants will be classified according to their chilling requirements to identify RAPD markers that cosegregate with chilling requirements. Presently, a bulked-segregant analysis is being performed on a tetraploid breeding population (primarily V. corymbosum) to identify RAPD markers linked to chilling requirement genes.

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Amnon Levi, Glenn A. Galau and Hazel Y. Wetzstein

A simple and efficient protocol is reported for the isolation of RNA from embryos and leaves of pecan [Carya illinoinensis (Wangenh.) K. Koch]. The method relies on suppression of the polyphenols from interaction with the RNA and their rapid removal from the homogenate by chloroform extraction. This method produced abundant amounts of high-quality RNA. This protocol is likely to be useful for Juglandaceous species and other recalcitrant plants with high levels of phenolic compounds.