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Bottle gourd [Lagenaria siceraria (Molina) Standl.] is widely produced in some Asian and African countries as a fresh vegetable as well as for seed consumption. A major use of bottle gourd is for rootstocks in grafted watermelon production. There are several centers where bottle gourd genetic resources are maintained, with the U.S. Department of Agriculture (USDA) germplasm collection being one of the most important. There is little published information on the relationship between seed morphology and the early establishment of seedlings in bottle gourd. The objective of this study was to determine seed characterization, morphology, and cotyledon shape in 163 Lagenaria spp. accessions and measure any relationship between seed and cotyledon characteristics. In USDA Lagenaria germplasm, it has been determined that the common character in seeds was medium in terms of seed size (53% of accessions), intermediate in seed surface lustre (39% of accessions), brown in seedcoat color (89% of accessions), thin and uniform in seed margin (35% of accessions), and tan in seed margin color (64% of accessions). According to the research results, seed weight ranged from 0.11 g (PI 500820) to 0.36 g (PI 675112), seed length from 13.17 mm (PI 500820) to 23.68 mm (PI 675112), and seed width from 5.86 (PI 500808) to 11.21 mm (PI 491274). Cotyledon length ranged from 5.46 cm (PI 368640) to 2.47 cm (PI 381850). The widest cotyledon was 3.00 cm (PI 534552), and the narrowest was 1.50 cm (PI 381831). Interesting correlations were observed for seed weight with seed length (R2 = 0.259), and cotyledon length with cotyledon width (R2 = 0.547).

Cucurbit plants usually are sensitive to chilling and easily damaged. Although bottle gourds, which are members of the Cucurbitaceae family, are considered as fresh vegetables in some Asian countries, their main use in recent years is to be used as rootstocks in grafted watermelon cultivation. We tested 163 bottle gourd accessions of the U.S. Department of Agriculture (USDA) genebank for cold tolerance in the early seedling stage. The experiment was conducted using controlled environment chambers with 3 chilling durations (36, 48, and 60 hours) at 4 °C. Chilling damage was rated 0 to 9 (0 = no damage, 1 to 2 = trace of damage, 3 to 4 = slight damage, 5 to 6 = moderate damage, 7 to 8 = advanced damage, 9 = plant totally dead). We rated damage separately for the cotyledons, true leaf, and growing point. Cold damage was higher at a chilling duration of 60 hours, and decreased at 48 and 36 hours. Most tolerant cultigens were PI 491272, PI 491280, PI 491281, PI 491286, and PI 491326. Most susceptible were PI 381845, PI 381846, PI 534556, PI 636137, and PI 668365.

Bottle gourd [Lagenaria siceraria (Molina) Standl.] landraces are widely grown in South Africa, and genetic diversity analysis is necessary to identify promising genotypes for breeding or systematic conservation. Sixty-seven diverse bottle gourd landraces were genotyped using 14 selected simple sequence repeat (SSR) markers. The number of alleles detected per marker ranged from 4 to 11, with a total of 86 putative alleles being amplified. Allele sizes ranged from 145 to 330 base pair (bp). Number of effective alleles (Ne) ranged from 1.58 to 6.14 with a mean of 3.10. Allelic richness varied from 3.00 to 8.90 with a mean of 5.23. Expected heterozygosity (He) values ranged from 0.37 to 0.84 with a mean of 0.65. The mean polymorphic information content (PIC) was 0.57. Jaccard’s coefficient of similarity values ranged from 0.00 to 1.00, with a mean of 0.63. Analysis of molecular variance (AMOVA) revealed that 79%, 17%, and 4% of the variation in bottle gourd landraces was attributable to among landraces, within landraces, and between populations, respectively. The study established the existence of considerable genetic diversity among South African bottle gourd landraces. Unique landraces such as BG-4, BG-6, BG-8, BG-9, and BG-15 from cluster I; BG-55, BG-42, BG-57, and BG-58 from cluster II; BG-28, BG-23, BG-29, and BG-34 from cluster III were selected based on their highest dissimilarity index. These could be useful for bottle gourd breeding and systematic conservation.

Powdery mildew (Podosphaera xanthii) can cause severe damage to cucurbit crops grown in open fields and greenhouses. In recent years, there has been an increased interest in the United States in grafting watermelon plants onto various cucurbit rootstocks. Bottle gourd plants (Lagenaria siceraria) are being used throughout the world as rootstocks for grafting watermelon. Although gourd plants are beneficial, they may still be susceptible hosts to various soilborne and foliar diseases. Bottle gourd plant introductions (PI) resistant to diseases and pests can be a valuable source of germplasm in rootstock breeding programs. We evaluated 234 U.S. PIs of L. siceraria for tolerance to powdery mildew in two greenhouse tests. Young seedlings were inoculated by dusting powdery mildew spores of melon race 1 on the cotyledons. Plants were rated 2 weeks after inoculation using a 1 to 9 scale of increasing disease severity. Although none of the L. siceraria PIs were immune to powdery mildew, several PIs had significantly lower levels of powdery mildew severity compared with susceptible watermelon cultivar Mickey Lee. The experiment was repeated with 26 select PIs on whole seedlings and cotyledon disks. Significant variability in the level of resistance to powdery mildew on plants within PI was observed. Moderate resistance in several PIs to powdery mildew was confirmed. PI 271353 had consistently lower ratings in the various tests and can be considered the most resistant to P. xanthii race 1 among the L. siceraria accessions evaluated in this study. A few other PIs with moderate resistance to powdery mildew included PI 271357, PI 381840, and PI 273663. These results suggest that novel sources of resistance could be developed by careful selection and screening of several of the PIs with moderate resistance described in our study.

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.

Confined-leaf tests in a greenhouse showed Lagenaria siceraria (Molina) Standley plant introduction (PI) 442369 was as susceptible to sweetpotato whitefly, Bemisia tabaci Gennadius, oviposition as Cucumis melo L., Cucurbita ecuadorensis Cutler and Whitaker, and Cucurbita lundelliana Bailey, whereas L. siceraria accessions PI 419090, PI 419215, PI 432341, and PI 432342 were resistant. Resistance rankings of L. siceraria accessions based on adult counts in greenhouse and field tests were similar. Adult entrapment among trichomes was highest on adaxial leaf surfaces of L. siceraria PI 419090. Abaxial leaf trichome density was 48.7/mm2 on sweetpotato whitefly-resistant L. siceraria PI 432342, 42.1/mm2 on Cucurbita lundelliana PI 540895, and ranged from 51.0 to 85.5/mm2 on Cucurbita ecuadorensis PI 540896. Leaf trichome densities of selected plants of four L. siceraria accessions ranged from 33.0 to 52/mm2 on the abaxial and from 6.3 to 20.8/mm2 on the adaxial surface. Scanning electron micrographs of the abaxial leaf surface, the preferred surface for oviposition, suggest that trichome configuration (density and arrangement of different lengths) could be a factor in reduction of whitefly oviposition on L. siceraria.

A procedure for producing transgenic bottle gourd plants by inoculating cotyledon explants with Agrobacterium tumefaciens strain AGL1 carrying a binary vector pCAMBIA3301, which contains glufosinate ammonium-resistant (bar) and the reporter (gus) genes, is describe. Infection was the most effective (highest infection frequency and index) when explants were co-cultivated with Agrobacterium for 6-8 days on co-cultivation medium supplemented with 0.001-0.1 mg/L L-a-(2-aminoethoxyvinyl) glycine (AVG). Transgenic plants were obtained with frequencies of about 0.2% when the explants were cultured on selection medium (MS medium supplemented with 3.0 mg/L BAP, 0.5 mg/L AgNO3, 500 mg/L cefotaxime, 2.0 mg/L DL-phosphinothricin, 0.3% sucrose and 0.8% Plant Agar. A histochemical gus assay, PCR and Southern blot analyses confirmed that transformation had occurred. Genetic analysis of T1 progenies showed that the transgenes were inherited in a Mendelian fashion. To our knowlege, this study represents the first report for Agrobacterium-mediated transformation in bottle gourd, rootstock for watermelon and other cucurbit crops in many countries.