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Fusarium wilt [caused by the fungus Fusarium oxysporum f. sp. niveum (FON)] has been a consistent problem in watermelon [Citrullus lanatus (Thunb.) Matsum. and Nakai] production worldwide. One method for combatting this pathogen in the field is to graft a susceptible, high-yielding scion on to a fusarium wilt-resistant rootstock. A concerning issue with rootstocks resistant to fusarium wilt is that they have not been tested for their susceptibility to plant pathogenic nematodes—specifically, root-knot nematodes (RKNs; Meloidogyne spp.) and the reniform nematode (Rotylenchulus reniformis). Preliminary findings have demonstrated that many of these Fusarium-resistant rootstocks are highly susceptible to RKNs. Research was conducted during the Spring and Fall 2015 and 2016 to evaluate the resistance to RKN and reniform nematode in rootstocks with known resistance to fusarium wilt. Six rootstocks were evaluated over the course of four experiments. A nematode-susceptible interspecific hybrid [Cucurbita maxima (Duchesne) × C. moschata (Duchesne)] rootstock ‘Carnivor’ was included as a susceptible control in both years. Results demonstrated that several Citrullus lanatus var. citroides (L.H. Bailey) rootstocks (‘Carolina Strongback’, USVL246-FR2, USVL252-FR2, and USVL-360) and ‘SP-6’ (a commercially available pollinizer cultivar) exhibited resistance to plant parasitic nematodes when compared with the susceptible control. Partial resistance was observed in USVL-482351. When compared with the control, these rootstocks also had fewer Meloidogyne spp. and R. reniformis in root tissue. These findings indicate that rootstocks may be available to manage both fusarium wilt and RKN in grafted cucurbit production system.

In this study, we report a simple procedure for developing and using new types of polymerase chain reaction (PCR) primers, named “high-frequency oligonucleotides–targeting active genes” (HFO-TAG). The HFO-TAG primers were constructed by first using a “practical extraction and report language” script to identify oligonucleotides (8, 9, and 10 bases) that exist in high frequency in 4700 expressed sequence tag (EST)-unigenes of watermelon (Citrullus lanatus) fruit. This computer-based screening yielded 3162 oligonucleotides that exist 32 to 335 times in the 4700 EST-unigenes. Of these, 192 HFO-TAG primers (found 51 to 269 times in the 4700 EST-unigenes) were used to amplify genomic DNA of four closely related watermelon cultivars (Allsweet, Crimson Sweet, Charleston Gray, and Dixielee). The average number of DNA fragments produced by a single HFO-TAG primer among these four watermelon cultivars was considerably higher (an average of 5.74 bands per primer) than the number of fragments produced by intersimple sequence repeat (ISSR) or randomly amplified polymorphic DNA (RAPD) primers (an average of 2.32 or 4.15 bands per primer, respectively). The HFO-TAG primers produced a higher number of polymorphic fragments (an average of 1.77 polymorphic fragments per primer) compared with the ISSR and RAPD primers (an average of 0.89 and 0.47 polymorphic fragments per primer, respectively). Amplification of genomic DNA from 12 watermelon cultivars and two U.S. Plant Introductions with the HFO-TAG primers produced a significantly higher number of fragments than RAPD primers. Also, in PCR experiments examining the ability of primers to amplify fragments from a watermelon cDNA library, the HFO-TAG primers produced considerably more fragments (an average of 6.44 fragments per primer) compared with ISSR and RAPD primers (an average of 3.59 and 2.49 fragments per primer, respectively). These results indicate that the HFO-TAG primers should be more effective than ISSR or RAPD primers in targeting active gene loci. The extensive EST database available for a large number of plant and animal species should be a useful source for developing HFO-TAG primers that can be used in genetic mapping and phylogenic studies of important crop plants and animal species.