Watermelon Cv Research Articles

First Report of Bacterial Wilt Caused by Erwinia tracheiphila on Pumpkin and Watermelon in New Mexico.

Pumpkin (Cucurbita pepo L., cv. Magic Lantern) and watermelon (Citrullus lanatus (Thunb.) Matsum. & Nakai, cvs. Millionaire and Sangrea) plants with wilting leaves and collapse of entire vines were observed during the 2005 and 2006 growing seasons in several fields in southwestern New Mexico (Luna and Hidalgo counties) with an incidence ranging from 7 to 25% and less than 1% in pumpkin and watermelon fields, respectively. Sticky, hyaline strands were visible when vines were cut transversally, indicative of bacterial wilt caused by Erwinia tracheiphila (2). In the pumpkin fields, 12-spotted cucumber beetles, vector insects of E. tracheiphila, were found on plants at the first-true-leaf stage, which were treated with dimethoate. At the 4- to 5-leaf stage, 5 to 10% of the plants were wilted and were removed by hand. Less than 1% of the plants showed symptoms prior to bloom, when a high population of beetles was observed, and the fields were treated with thiamethoxam. To isolate the causal agent of the wilt symptoms, six, 1-cm vine segments and three to five beetles were surface sterilized in 1% NaOCl for 2 min, rinsed and macerated in sterile distilled water, and plated onto potato dextrose agar, nutrient agar, and King's medium B. After incubation at 25°C, bacterial colonies emerged on all media and were grayish white-to-cream, circular, smooth, and glittering. Isolated bacteria were gram negative, did not grow at 39°C, produced hydrogen sulfide gas from hydrolysis of cysteine, and did not hydrolyze litmus milk and starch. With Ready-To-Go PCR beads and 16S rDNA-based primers ET1/ET2 (1), a 700-bp product was obtained from each of two isolates, consistent with previously reported data for E. tracheiphila (1,3). For the pathogenicity tests, 10 seedlings of pumpkin cv. Magic Lantern and watermelon cv. Millionaire were inoculated with each isolate in the greenhouse at the second fully expanded leaf stage using two methods. In the first method, stems were injected with bacterial suspension (106 CFU/ml) using a hypodermic needle. In the second method, a dab of bacterial colonies was taken with a sterile toothpick to stab the cotyledonary axils. Control seedlings were stem injected with distilled water or stabbed with a sterile toothpick. The experiments were conducted four times. Inoculated plants were placed in a humid chamber at 23 to 25°C. Plant wilting was observed within 4 days when stab inoculated with toothpicks and within 7 to 10 days when stem injected with bacterial suspension. Bacterial colonies recovered from inoculated plants were identical to those recovered from field infected plants. To our knowledge, this is the first report of bacterial wilt on pumpkin and watermelon in New Mexico.

English

Watermelon (Citrullus lanatus) cultivars are highly susceptible to the southern root-knot nematode (Meloidogyne incognita), with considerable yield losses when this nematode is not managed. Empirical evidence suggested that wild watermelon (Cucumis africanus) and wild cucumber (Cucumis myriocarpus) were highly resistant to M. incognita race 2. The objective of this study was two-fold; viz. to determine whether (1) C. africanus and C. myriocarpus seedling rootstocks would be compatible with watermelon cv. ‘Congo’ and ‘Charleston Gray’ and (2) the twoCucumis spp. rootstocks would retain their nematode-resistance capabilities when grafted with the two highly nematode-susceptible watermelon cultivars. The eight treatment combinations were arranged in a randomised complete block design, with six replications. At harvest, 56 days after transplanting the grafted seedlings, with highly susceptible watermelon cultivars had no effect on nematode-resistance capabilities of C. africanus and C. myriocarpus. Also, the two Cucumis spp. were compatible with the two watermelon cultivars. Consequently, C. africanus and C. myriocarpus rootstocks have the potential for use as resistant rootstocks in the management of M. incognita race 2 in watermelon production.   Key words: Cucumis spp., root galls, resistant rootstocks, reproductive factor, watermelon cultivars.

LATEST ADVANCES IN WATERMELON GENOMICS

Watermelon is an important vegetable crop throughout the world and its worldwide harvested area is 21.7% of that of all vegetables. The genetic diversity of cultivated watermelon is very narrow, which is a major limiting factor of its breeding. Therefore it is necessary to develop new resources that can be used to widen watermelon genetic basis. We have initiated the Watermelon Whole Genome Sequencing project and currently have generated Solexa sequences which represent more than 89.9X coverage of the genome of watermelon cv. '97103' and covered 95% of the whole genome and 98% of the genic region. Further genome sequencing of '97103' using the traditional Sanger technology, comparative genome sequencing of watermelon cv. '90-4304' (10X coverage Solexa sequences), genome annotation is currently underway. In addition, using Roche 454 GS-FLX massively parallel pyrosequencing technology, we have generated 715,059 ESTs with an average length of 250 bp from fruits of '97103' during different developmental stages. These sequences are being assembled and functionally annotated. SSR markers and pathway prediction have been analyzed preliminarily. A comprehensive digital expression map during watermelon fruit development is being created. Furthermore, we have constructed a SSH library from root tissues of watermelon cv. 'PI296341-FR' infected by Fusarium wilt racel and randomly sequenced approximately 3,895 SSH clones. Sequence analysis indicated that 28.3% of the unique ESTs were related to defense response, signal transduction and interaction with the environment. GFP gene is used as the marker to detect the infection process of Fusarium wilt in root tissue. Three hundred twenty-four SSR and EST-SSR markers consist of the watermelon RIL SSR genetic map. The relationships between four Chromosomes and the genome are identified by FISH.

First Report of Monosporascus cannonballus on Watermelon in Brazil.

In 2008 and 2009, vine decline symptoms were observed in three watermelon (Citrullus lanatus (Thunb.) Matsum. & Nakai) fields located in the municipalities of Mossoró (Rio Grande do Norte State) and Quixeré (Ceará State) in northeastern Brazil. Symptoms included yellowing of crown leaves just prior to harvest and collapse of many of the vines. Mean maximum daily temperatures for the first and second half of the season were 28.6 and 25.1°C, respectively. Affected plants exhibited necrotic root systems and lacked most of the secondary and tertiary feeder roots. Numerous perithecia on the roots contained asci and ascospores characteristics of Monosporascus cannonballus Pollack & Uecker (1,2). Small pieces of primary and secondary roots were surface disinfected and plated onto potato dextrose agar (PDA) medium with 0.5 g liter-1 of streptomycin sulfate and incubated for 7 days at 25°C in the dark. Hyphal tips from all colonies were transferred to PDA and further incubated for 30 to 40 days at 25°C in the dark for subsequent growth and sporulation. Isolations consistently yielded colonies of white mycelium, which became dark grayish after 10 to 15 days, and perithecia with one-spored asci. The internal transcribed spacer regions of ribosomal DNA of isolates 18-5 and 19-1 were sequenced (GenBank Accession Nos. GQ891544 and GQ891545). These sequences were identical to sequences of M. cannonballus (GenBank Accession Nos. AM167936 and AM167937). Pathogenicity of these two isolates was confirmed on watermelon cv. Crimson Sweet in a greenhouse maintained at 25 to 30°C. Inoculum was produced in a sand-oat hulls (Avena sativa) medium (0.5 liter of sand, 46 g of ground oat hulls, and 37.5 ml of distilled water) and incubated at 25°C for 1 month. CFU were quantified by serial dilution using 1% hydroxyethyl cellulose. A sterilized mixture of equal portions (vol/vol) of sand and peat moss was used to fill 17-cm-diameter plastic pots and inoculum was added to produce an inoculum concentration of 20 CFU g-1. Five watermelon seeds planted in each pot were later thinned to one seedling per pot. There were five replicated pots for each treatment with an equal number of noninfested pots. Plants were evaluated for disease 45 days after sowing. All isolates of M. cannonballus were highly aggressive and caused severe root necrosis compared with the noninoculated controls. M. cannonballus was reisolated from symptomatic plants, confirming Koch's postulates. In 2004, M. cannonballus was reported in the same Brazilian cucurbit-growing areas causing root rot and vine decline of muskmelon (Cucumis melo L.) (3), but to our knowledge, this is the first report of M. cannonballus on watermelon in Brazil.

First Occurrence of Cucurbit Powdery Mildew Caused by Race 3-5 of Podosphaera fusca in Spain.

A new race of cucurbit powdery mildew was observed for the first time on melon (Cucumis melo) in three research greenhouses in the Axarquia area of southern Spain during the spring of 2008. Fungal growth appeared as white powdery colonies initially restricted to the upper leaf surfaces. Morphological characteristics of colonies, conidiophores, conidia, germ tubes, and appressoria indicated that the powdery mildew fungus was Podosphaera fusca (also known as P. xanthii) (3), a fungal pathogen extensively reported in the area (1). However, the fungus developed on plants of melon cv. PMR 6, which is resistant to races 1 and 2 of P. fusca, suggesting that the fungus could belong to race 3, a race of P. fusca not yet reported in Spain. Race determination was carried out by inoculating the third true leaf of a set of differential melon genotypes that were maintained in a greenhouse. Symptoms and colonization observed on cvs. Rochet, PMR 45, PMR 6, and Edisto 47 indicated that the isolates belonged to race 3-5 of P. fusca. Fungal strains of races 1, 2, and 5 of P. fusca (all present in Spain) were used as controls. Pathotype designation was determined by inoculating different cucurbit genera and species (2). In addition to melon, the isolates were pathogenic on zucchini (Cucurbita pepo) cv. Diamant F1, but failed to infect cucumber (C. sativus) cv. Marketer and watermelon (Citrullus lanatus) cv. Sugar Baby; therefore, the isolates were pathotype BC (2). Races 1, 2, 4, and 5 of P. fusca have been previously reported in the area (1). The occurrence of race 3-5 represents another challenge in the management of cucurbit powdery mildew in Spain.

First European Report of a Leaf Spot on Common Ragweed (Ambrosia artemisiifolia) Caused by a Phoma sp.

Common ragweed, native to North America, has recently become invasive in some parts of Europe. In Hungary, it has become the most widespread agricultural weed species and the most important producer of allergenic pollen since the 1990s. During surveys for its fungal plant pathogens to be evaluated as potential biological control agents (1), ragweed plants exhibiting necrotic spots on the leaves and stems were repeatedly found in Heves and Vas counties in Hungary in September 2004 and 2006. Numerous globose and ostiolate pycnidia, 68 to 115 μm in diameter, containing hyaline, unicellular conidia, 3 to 8 μm long, were found in necrotic tissues. On the basis of these characteristics, the fungus was identified as a Phoma sp., and 21 isolates were obtained on Czapek-Dox medium supplemented with 2% malt and 0.5% tetracycline in 2004 and 2006. Two well-sporulating isolates, designated Ph-5 and Ph-17, were selected for further studies. DNA was extracted from mycelium with a Qiagen DNeasy Plant Kit (Hilden, Germany) and the rDNA internal transcribed spacer (ITS) sequences were amplified and determined as described by Szentiványi et al. (2). The ITS sequences were identical in these two isolates and were 97 to 98% similar to those of Didymella bryoniae (anamorph Phoma cucurbitacearum), a pathogen of cucurbits, and also to those of other Phoma spp. No ITS sequences identical to those determined in Phoma isolates Ph-5 and Ph-17 were found in GenBank. Sequence data were deposited in GenBank (No. FJ794609). To test the pathogenicity of Ph-17 grown on Czapek-Dox medium with 2% malt, a 2 to 6 × 105 conidia/ml aqueous suspension was used to inoculate 2-month-old potted ragweed plants and 1-month-old cucumber cv. Rajnai fürtös, bottle gourd (Lagenaria leucantha) cv. Minibottle, and watermelon (Citrullus lanatus) cv. Sugar Baby, which were all grown from seeds in a greenhouse. Plants were kept in transparent plastic chambers for 6 weeks. Five pots with one to three plants each were used for each plant species tested and the experiment was carried out twice. Noninoculated plants, two pots with one to three individuals for each species kept in the same way, served as controls. Necrotic spots with pycnidia developed on 38 to 47% of the leaves of all inoculated ragweed plants 18 to 25 days after inoculation, whereas all the cucurbitaceous plants tested, as well as the control ragweed plants, did not develop disease symptoms. Although the Phoma isolate Ph-17 was, based on ITS sequence data, closely related to D. bryoniae, it was not pathogenic to cucurbits. The pathogen was reisolated from two diseased ragweed plants. Several Phoma spp. strains were isolated from Ambrosia artemisiifolia in the United States and Canada (3,4), but to our knowledge, none were isolated outside North America. One of the strains has already been used as a potential biological control agent of ragweed in Canada, but then lost its virulence in culture (3). The biocontrol potential of the Hungarian Phoma sp. isolate Ph-17 against A. artemisiifolia is currently being investigated. To our knowledge, this is the first report of a Phoma sp. on A. artemisiifolia in Europe.

First Report of Xanthomonas cucurbitae Causing Bacterial Leaf Spot of Watermelon in the Seychelles.

Bacterial leaf spot of cucurbits caused by Xanthomonas cucurbitae (4) can be a harmful disease of several cucurbit species in tropical environments, mainly within the Cucumis, Cucurbita, and Citrullus genera. The bacterium induces angular, water-soaked leaf spots, which sometimes become necrotic and have a chlorotic halo. Scab-like lesions on fruit can also be observed (2). Water-soaked, angular leaf lesions were collected from approximately 15 watermelon plants (Citrullus lanatus) in a production field located in Mahé, Seychelles in 2003. Yellow-pigmented Xanthomonas-like bacterial colonies were isolated on KC semiselective medium (yeast extract 7 g, peptone 7 g, glucose 7 g, agar 18 g, distilled water 1,000 ml, propiconazole 20 μg ml-1, cephalexin 40 mg liter-1, and kasugamycin 20 mg liter-1) from all isolation attempts (3). Amplified fragment length polymorphism (AFLP) analysis was performed on four watermelon strains together with reference strains of Xanthomonas cucurbitae (LMG 690 [type strain] and LMG 8663) and the type strain of all other valid Xanthomonas species using SacI/MspI and four primer pairs (unlabeled MspI + 1 [A, C, T, or G] primers and 5'-labeled - SacI + C primer for the selective amplification step) (1). The four strains from watermelon showed identical fingerprints and were most closely related to X. cucurbitae. One strain from diseased watermelon (JZ88-1) was further analyzed by MultiLocus Sequence Analysis (MLSA) using portions of three housekeeping genes (atpD, dnaK, and gyrB) as described previously (1). This strain displayed a very high relatedness (99.8 and 98.9% with strain LMG 690 and LMG 8663, respectively) to the two reference strains of X. cucurbitae. AFLP and MLSA were useful for identifying strains at the species level that were consistent with previous results (1). Bottle-gourd (Lagenaria siceraria), pumpkin (Cucurbita maxima), squash cv. Aurore (Cucurbita pepo), cucumber cv. L-04 (Cucumis sativus), cantaloupe melon cv. Cezanne (Cucumis melo), and watermelon cv. Fou-nan (C. lanatus) leaves were infiltrated (10 inoculation sites per leaf and three replicates) with bacterial suspensions (JZ88-1, LMG 690 and LMG 8663) containing approximately 1 × 105 CFU ml-1 (approximately 1 × 102 CFU per inoculation site). Typical water-soaked lesions that developed into necrotic spots were observed 6 to 8 days after inoculation for all inoculated strains on all inoculated plant species. One month after inoculation, Xanthomonas was recovered from lesions and population sizes determined on KC semiselective medium (3) ranging from 1 × 106 to 9 × 106 CFU per lesion were typical of a compatible interaction. Bacterial leaf spot has appeared sporadically in Mahé, Seychelles since 2003, most often with limited incidence. However, growers need to be aware of the potential negative effect of this disease in tropical environments.

First Report of Xanthomonas cucurbitae Causing Bacterial Leaf Spot of Pumpkin on Réunion Island.

Bacterial leaf spot of cucurbits caused by Xanthomonas cucurbitae (3) can be of economic importance in tropical and subtropical production areas, most often within the Cucumis, Cucurbita, and Citrullus genera. The bacterium induces angular, water-soaked leaf spots, which sometimes turn necrotic with a chlorotic halo. Scab-like lesions on fruit may also be observed (1). During 2000, water-soaked, angular leaf lesions were collected from pumpkin (Cucurbita pepo) in a production field located at Petit Serré, Réunion Island. Yellow-pigmented Xanthomonas-like bacterial colonies were isolated on yeast peptone glucose agar. Amplified fragment length polymorphism analysis was performed on four pumpkin isolates together with reference strains of X. cucurbitae (LMG 690 [type strain] and LMG 8663) and the type strain of all other valid Xanthomonas species using SacI/MspI and four primer pairs (unlabeled MspI + 1 [A, C, T, or G] primers and 5'-labeled - SacI + C primer for the selective amplification step) (N. Ah-You, L. Gagnevin, P. A. D. Grimont, S. Brisse, X. Nesme, F. Chiroleu, L. Bui Thi Ngoc, E. Jouen, P. Lefeuvre, C. Verniére, and O. Pruvost, personal communication). The four isolates from pumpkin showed identical fingerprints and were most closely related to X. cucurbitae, with evolutionary genome divergences ≤0.05 (N. Ah-You et al., personal communication). One strain from diseased pumpkin (JW210-1) was further analyzed by multilocus sequence analysis using three housekeeping gene portions (atpD, dnaK, and gyrB) as described previously (N. Ah-You et al., personal communication). Although not fully identical, this strain displayed a similarity of >99% to the two reference strains of X. cucurbitae. Pumpkin and bottle-gourd (C. maxima), squash cv. aurore (C. pepo), cucumber cv. L-04 (Cucumis sativus), melon cv. cezanne (Cucumis melo), and watermelon cv. fou-nan (Citrullus lanatus) leaves were infiltrated (10 inoculation sites per leaf; three replicates) with bacterial suspensions prepared from strains JW210-1, LMG 690, and LMG 8663 and containing approximately 1 × 105 CFU ml-1. Negative controls consisted of leaves infiltrated with sterile tris buffer. Typical, water-soaked lesions that developed into necrotic spots were observed 6 to 8 days after inoculation for all inoculated plant species-strain combinations, but not for negative controls. One month after inoculation, mean Xanthomonas population sizes recovered from leaf lesions on KC semiselective medium (2) ranged from 1 × 107 to 1 × 108 CFU per lesion, typical of a compatible interaction. The reported outbreak was circumscribed to a single field and did not affect the local industry. No major outbreak of bacterial leaf spot of cucurbits has been reported on Réunion Island since 2000 on any host species of X. cucurbitae.

Effect of Incorporation of Brassica spp. Residues on Population Densities of Soilborne Microorganisms and on Damping-off and Fusarium Wilt of Watermelon.

Incorporating Brassica spp. residue to reduce populations of soilborne fungi and manage damping-off and Fusarium wilt of watermelon (Citrullus lanatus var. lanatus) was studied in two field experiments. Treatments included incorporating flowering Brassica napus cv. Dwarf Essex canola or B. juncea cv. Cutlass mustard and laying black polyethylene mulch at incorporation or 1 month after incorporation, methyl bromide, and a nontreated control. In both years, glucosinolates were identified and quantified in the shoots and roots of the flowering plants. In both years, the total concentration of glucosinolates incorporated per square meter was significantly higher for B. juncea than for B. napus. Isothiocyanates were inconsistently detected in the amended soils and none were detected more than 12 days postincorporation. After incorporation in 2004 and 2005, amended plots had higher populations of Fusarium oxysporum and Pythium spp. than the methyl bromide treatment, and in some treatments, populations were higher than in the control. Fluorescent Pseudomonas spp. were not suppressed in amended soils, and their populations were significantly higher in some amended treatments than those in methyl bromide-treated soils or nontreated control soils. Incidence of damping-off and severity of Fusarium wilt on seedless watermelon cv. Tri-X 313, which is susceptible to Fusarium wilt, were not consistently lower in brassica-amended soils or methyl bromide-treated plots than in nontreated control plots. Therefore, under spring conditions and methods used in this study, neither biofumigation nor methyl bromide fumigation in coastal South Carolina was an effective disease management tool for two soilborne pathogens of watermelon.

First Report of Acidovorax avenae subsp. citrulli Infecting Edible Seed Watermelon (Citrullus lanatus var. lanatus) in China.

Edible seed watermelon, Citrullus lanatus var. lanatus, is a type of watermelon from which only the seeds are consumed. Between 2003 and 2005, a bacterial disease was discovered on this type of watermelon in several regions of Xinjiang Province in western China, with a diseased area of 40,000 ha. The average disease incidence on fruit and seedlings was approximately 30 and 20%, respectively. Symptoms were most noticeable on fruit. Lesions on the fruit rind were first noticeable as small pinpoint water-soaked areas. As the lesions rapidly enlarged, black, starshaped cracks formed in the rind, and the rind along with the flesh around the spots hardened. If no secondary invaders or saprophytes entered through the cracks, the inside flesh often disintegrated into dry stiff cavities. The fruit with necrotic spots became markedly malformed. The pathogen also infected seedlings. First symptoms on the seedling appeared as dark, water-soaked lesions on the underside and edge of the cotyledons. As the cotyledon expanded, the lesions become necrotic and often extended along the length of the midrib and eventually merged into large, black, withered areas. To determine the causal agent of the disease, 54 bacterial strains were isolated from cotyledons, fruits, or seeds of edible seed watermelon. By spray inoculation, these strains were inoculated onto edible seed watermelon cv. Xinzigua1 at the two-true-leaf stage with bacterial concentrations at 107 CFU/ml. All strains were strongly virulent to cv. Xinzigua1, which is planted in large areas in Xinjiang, by forming many water-soaked spots on cotyledons and true leaves. There were no symptoms in the control that was sprayed with sterile deionized water. Through biological and biochemical characteristics, including Gram reaction and tests for catalase, oxidase, and oxygen requirement, all strains were identified as Acidovorax avenae subsp. citrulli. To verify pathogen identification, specific polymerase chain reaction was also carried out. One set of 16S primers, WFB1/WFB2 for A. avenae subsp. citrulli, was used (1). A single unique band of approximately 360 bp was amplified for all strains tested. To our knowledge, this is the first report of A. avenae subsp. citrulli infecting edible seed watermelon. Reference: (1) R. R. Walcott and R. D. Gitaitis. Plant Dis.84:470, 2000.

Occurrence of Bacterial Fruit Blotch of Watermelon Caused by Acidovorax avenae subsp. citrulli in the Eastern Mediterranean Region of Turkey.

Turkey has the second highest production level of watermelon (Citrullus lanatus L.) in the world, with 3.8 million tons produced in 2005. In the spring of 2005, a severe outbreak of a disease resembling bacterial fruit blotch was observed on watermelon cv. Crispy growing in a production area of 35,000 ha in Adana Province in the eastern Mediterranean Region of Turkey. In 13 commercial watermelon fields surveyed in this study, incidence of symptomatic fruit ranged from 30 to 45%. Characteristic symptoms were dark, gray-green stains or blotches on the surface of fruits. Cracks developed in the rind and released an amber-colored substance. Bacteria were consistently isolated from affected fruits and they formed nonfluorescent colonies on King's medium B. Identification of isolated bacterial strains was based on the methods described by Schaad et al. (2). All strains were oxidase positive, gram negative, arginine dyhydrolase negative, and produced a hypersensitive response on tobacco leaves (Nicotiana tabacum cv. Samsun). Strains were identified as Acidovorax avenae subsp. citrulli (1,2) on the basis of the results of biochemical tests (1,2) and sole carbon substrate utilization (BIOLOG GN; Biolog Inc., Hayward, CA) with 75 to 93% similarity indices. A pathogenicity test was performed for nine identified strains by injecting a bacterial suspension (108 CFU/ml of saline) under the rind of three harvested watermelon (cv. Diyarbakir) fruits and into the stems of three seedlings. Saline was used as the negative control. The fruits and seedlings inoculated with each strain were incubated in polyethylene bags for 48 h at 25°C. Dark green water-soaked lesions developed on inoculated fruits and water-soaking and stem necrosis were observed on the seedlings 3 to 5 days after inoculation. The bacterium was readily reisolated from inoculated fruits and seedlings and identified as A. avenae subsp. citruli on the basis of BIOLOG GN. In 1995, bacterial fruit blotch of watermelon has been observed and eradicated in the Marmara Region of western Turkey (1). Therefore, to our knowledge, this is the first outbreak of bacterial fruit blotch on watermelon in the eastern Mediterranean Region of Turkey.

First Report of a Leaf Spot and Stem Canker Caused by Myrothecium roridum on Watermelon in the United States.

Myrothecium roridum Tode:Fr, pathogenic to a number of cucurbit species, causes fruit rots, cankers on crowns and stems, and leaf spots. Hosts include cantaloupe and honeydew (Cucurbita melo) and cucumber (Cucumis sativus) (1,3). In June 2004, following a period of heavy rainfall, numerous round-to-oblong, brown lesions with concentric rings were observed on leaves of watermelon (Citrullus lanatus) cv. Desert King at the Blackshank Farm in Tifton, GA. Disease was localized in the field and severity was low (<5% of leaf area affected). No symptoms were observed on fruit. Sections of tissue were removed from the margin between healthy and diseased tissue and plated on acidified, 25% potato dextrose agar (aPDA). A small plug of agar and mycelium were removed from colonies that emerged from lesions and were transferred to aPDA. Isolated colonies were characterized by a white, floccose mycelium with concentric, dark green-to-black rings of sporodochia bearing viscid masses of conidia. Conidia were cylindrical with rounded ends and measured 6 to 8 × 1.5 to 2.5 μm. The features of the fungus were consistent with the description of Myrothecium roridum (1,2). Pathogenicity tests were conducted in a temperature-controlled greenhouse. Twenty-five watermelon plants (cv. Desert King) were inoculated with a conidial suspension of M. roridum (5 × 105 conidia per ml) plus 0.1% vol/vol Tween 20. Inoculum was applied on leaves and stems until runoff with a hand-held mister, and plants were placed in a dew chamber for 72 h. Ten plants were sprayed with sterile, distilled water to serve as controls. Inoculated and noninoculated control plants were removed from the dew chamber and maintained at 25 to 28°C. Symptoms appeared 8 days after inoculation and were characterized by round, dark lesions with concentric rings; noninoculated plants were symptomless. Sections of symptomatic tissue were plated, and M. roridum was reisolated. Although M. roridum is a common pathogen of melons and cucumber, to our knowledge, this is the first field report of a leaf spot caused by M. roridum on watermelon in the United States. No further occurrences of the disease on watermelon have been observed in Georgia since the initial discovery of M. roridum in 2004; however, losses could be potentially severe if widespread infection of fruit were to occur.

Response of cucurbit species to Rotylenchulus reniformis

The state of Rio Grande do Norte is the major melon (Cucumis melo) producer in Brazil and the reniform nematode Rotylenchulus reniformis has become increasingly important due to damages to that crop and the lack of resistant cultivars and effective control measures. Under these circumstances, eight cucurbit genotypes were screened for resistance or tolerance to a population of R. reniformis in naturally infested field soil, under greenhouse conditions. A high reduction in shoot mass was found in all infected genotypes. Watermelon (Citrullus lanatus) cv. Sugar Baby showed the lowest rate of nematode reproduction.

Activation of Isopentenyl Transfarese (ipt) Gene in Transformed Watermelon Influences Seed Number and Other Characteristics

The watermelon cv. Crimson Sweet was transformed with the copper inducible isopentenyl transferase, the rate-limiting step in cytokinin biosynthesis, gene via Agrobacterium tumafaciences (LBA4404). Transformed (ipt) and nontransformed plants were regenerated from tissue culture and clonally propagated by the rooting of leaf node cuttings. Twelve plants of each were grown in 1-gal. pots. Once the plants initiated new growth both transgenic plants and wild type plants were sprayed with one of four different concentrations (0, 5, 10, &amp; 50 μm) of CuSO4. The experimental unit was a single plant with three replicates. The growth rate, number of leaves, flowers, lateral shoots, and chlorophyll content were measured weekly for five weeks. Treated transgenic plants had greater numbers of leaves, flowers and lateral branches as well as higher chlorophyll levels. Pollen viability was examined in all treatments with no differences among treatments. Plants of both types were self pollinated to generate seeds. Female flowers were bagged before opening and then selfed. Selfed flowers were bagged for at least two days. The fruits were grown for eight to ten weeks with support. Once they reached maturity, fruits were harvested and fruit shape, flesh color, brix, number of normal seeds, number of colored but empty seeds and number of white seeds were recorded. Significant differences were observed only in seed number between wild type and transgenic (both treated and nontreated,) watermelon fruits. The number of seeds in transgenic watermelon plants treated with CuSO4 was reduced to about 5% to 7% of wild type plants. Transgenic plants which received no CuSO4 had approximately 33% to 50% of the seed of wild type.

Evaluation of Resistance to Gummy Stem Blight (Didymella bryoniae) Disease in ipt (Isopentenyl Transferase) Transformed Watermelon Plants

Watermelon plants are susceptible to Gummy stem blight disease that considerably reduces yields worldwide. In order to develop non-specific resistance, watermelon cv. Crimson Sweet was transformed with copper inducible isopentenyl transferase (ipt), the rate-limiting step in cytokinin biosynthesis, gene via Agrobacterium tumafaciences (LBA4404). Transformed (ipt) and nontransformed plants were grown at approximately 28-30 °C day, 20-22 °C night and 16 hours daylight under greenhouse conditions. Once the plants initiated new growth both transgenic plants and wild type plants were sprayed with one of three different concentrations (0, 10 &amp; 50 μm) of CuSO4. Plants were sprayed twice to run-off in a twenty-four hour time period before inoculation with the pathogen. Cultures of the pathogen Didymela bryonia (W353) were grown for about 3 weeks and an inoculum containing 105 conidia per mL was sprayed with the prepared suspension until initial run-off in a humidified chamber. The disease symptoms were evaluated after one week with resistance demonstrated in all treated transgenic plants. All nonsprayed transgenic and wild type plants showed similar disease symptoms. Infected leaf samples were surface sterilized and re-cultured on V8 medium. The characteristics of the recovered pathogen confirmed that it was identical to the stock culture of W353. The same experiment has been conducted on seedlings from transgenic (T1 generations) and non-transformed plants. The non-transformed seedlings showed the first disease symptoms on their cotyledons and lower leaves. Disease resistance was observed in seedlings of the treated transformed plants as compared to nontransformed ones.

Differences in Pathogenicity between two Genetically Distinct Groups of Acidovorax avenae subsp. citrulli on Cucurbit Hosts

AbstractUsing DNA fingerprinting by pulse‐field gel electrophoresis and repetitive extragenic pallindromic (REP)‐polymerase chain reaction (PCR), two distinct groups were confirmed among 64 Acidovorax avenae subsp. citrulli strains collected from a range of cucurbitaceous hosts in the USA, China, Taiwan, Thailand, Canada, Australia, Brazil and Israel. Eighty‐two percent of the group I strains were recovered from non‐watermelon hosts and the subspecies type strain was the only member of this group that utilized l‐leucine as a sole carbon source. On the contrary, 94% of the group II strains were recovered from watermelon and 96% of them utilized l‐leucine. Two‐week‐old watermelon cv. Crimson sweet, cantaloupe cv. Athena, pumpkin cv. Lumina and squash cv. Early yellow crookneck seedlings were susceptible to A. avenae subsp. citrulli strains representing each group with the exception of the subspecies type strain. Overall, seedlings of watermelon cv. Crimson Sweet were most susceptible to A. avenae subsp. citrulli infection followed by cantaloupe, pumpkin and squash. Group II strains were more aggressive watermelon than on other hosts. On the contrary, group I strains were moderately aggressive on all cucurbit hosts tested.

In vitro Decline in Plant Cultures: Detection of a Legion of Covert Bacteria as the Cause for Degeneration of Long-term Micropropagated Triploid Watermelon Cultures

In vitro cultures of triploid seedless watermelon cv. Arka Manik displayed a decline in shoot and root growth after 4–5 years of active culturing. Visibly clean cultures upon indexing on enriched media showed covert bacteria, and a significant improvement in proliferation and rooting in response to surface sterilization. The bacteria however survived endophytically. Low pH and reduced clarity of agar gelled medium were found to mask the expression of bacteria in the tissue culture medium. Gentamycin, streptomycin or broad-spectrum bactericide cefazolin provided as a 2 ml overlay in the medium in factorial combination at 0 or 50 mg l−1 resulted in selective suppression of some bacteria depending on the treatment and eight bacterial clones comprising of four Gram-positive (Bacillus spp.) and four-Gram negative (3 × Pseudomonas spp. and 1 × Aeromonas sp.) strains were isolated from the cultures. Provision of 50 mg l−1 gentamycin in 2 ml overlay in the multiplication or rooting medium coupled with occasional decontamination of cultures helped in circumventing the decline problem. The plants established in the field after 6 years of active in vitro culturing appeared normal and fertile suggesting the feasibility of keeping cultures for long periods, thus saving time and other resources. Freeing the cultures from covert bacteria was complicated by the presence of different bacterial types and this will be addressed later.

Races and Inoculum Density of Fusarium oxysporum f. sp. niveum in Commercial Watermelon Fields in Maryland and Delaware.

A survey was conducted to determine races and inoculum density of Fusarium oxysporum f. sp. niveum, the causal agent of Fusarium wilt of watermelon in Maryland and Delaware. Virulence on six differential cultivars was tested for each of 63 isolates of F. oxysporum f. sp. niveum, obtained from 25 commercial watermelon fields. Thirteen isolates (21%) were identified as race 0, 36 isolates (57%) as race 1, and 14 isolates (22%) as race 2. Races 0 and 1 were present in 12 (48%) and 10 (40%) of the fields, respectively. The highly aggressive race 2 was identified from five fields in two counties in Maryland and from one field in Delaware, representing 24% of the fields. Race 2 was copresent with one or two other races. Race 2 (19 isolates) predominated among the 25 isolates obtained from a research field in Maryland. Nineteen commercial fields had inoculum densities of F. oxysporum f. sp. niveum ranging from 100 to 1,200 CFU/g of soil at harvest. Within this range of inoculum densities, >20% incidence of wilt was observed when the susceptible watermelon cv. Sugar Baby was planted in samples of soil collected from these fields. The relationship (P < 0.0001) between inoculum density of F. oxysporum f. sp. niveum (X) and incidence of Fusarium wilt (Y) on Sugar Baby was best described using the monomolecular equation, Y = 1 - exp[-0.0013 (X + 166)]. The ratio of pathogenic to total population of F. oxysporum in the fields linearly increased with increasing inoculum density of F. oxysporum f. sp. niveum (R 2 = 0.4; P < 0.0009).

Rootstock resistance to Fusarium wilt and effect on watermelon fruit yield and quality

The potential of grafted watermelon for resistance toFusarium oxysporum f.sp.niveum on some Curcurbitaceae,Lagenaria, Luffa, Benincasa and commercial rootstocks was evaluated. Effects of grafting on yield and quality of diseased plants were evaluated. All grafted plants and rootstocks were resistant to the three known races (0, 1, and 2) ofF. oxysporum f.sp.niveum except watermelon cv. ‘Crimson Tide’, which was susceptible to race 2. Fruit yield was positively (21–112%) affected byLagenaria rootstocks but negatively affected (200–267%) byCucurbita rootstocks when compared with the control. While only minor differences in fruit quality were determined in control and grafted plants onLagenaria rootstocks, the quality parameters for watermelon grafted ontoCucurbita rootstocks were lower than in the control. The reasons for low yield and quality might be due to an incompatibility betweenCucurbita rootstocks and watermelon. These results showed that rootstock influence on disease resistance as well as yield and quality of scion fruit is important in determining the potential use of grafting applications in watermelon.

Mycorrhizal colonisation improves fruit yield and water use efficiency in watermelon (Citrullus lanatus Thunb.) grown under well-watered and water-stressed conditions

The effect of arbuscular mycorrhizal (AM) colonisation by Glomus clarum on fruit yield and water use efficiency (WUE) was evaluated in watermelon (Citrullus lanatus) cv. Crimson Sweet F1 under field conditions. Treatments were: (1) well-watered plants without mycorrhizae (WW-M), (2) well-watered plants with mycorrhizae (WW+M), (3) water- stressed plants without mycorrhizae (WS-M) and (4) water-stressed plants with mycorrhizae (WS+M). When soil water tension readings reached −20 and −50 kPa for well-watered (WW) and water-stressed (WS) treatments, respectively, irrigation was initiated to restore the top soil to near field capacity. Water stress reduced watermelon shoot and root dry matter, fruit yield, water use efficiency but not total soluble solids (TSS) in the fruit, compared with the non-stressed treatments. Mycorrhizal plants had significantly higher biomass and fruit yield compared to nonmycorrhizal plants, whether plants were water stressed or not. AM colonisation increased WUE in both WW and WS plants. Macro- (N, P, K, Ca and Mg) and micro- (Zn, Fe and Mn) nutrient concentrations in the leaves were significantly reduced by water stress. Mycorrhizal colonisation of WS plants restored leaf nutrient concentrations to levels in WW plants in most cases. This is the first report of the mitigation of the adverse effect of water stress on yield and quality of a fruit crop.