Adhikari, Mahesh;Yadav, Dil Raj;Kim, Sang Woo;Um, Young Hyun;Kim, Hyun Seung;Lee, Seong Chan;Song, Jeong Young;Kim, Hong Gi;Lee, Youn Su
The Plant Pathology Journal
/
v.33
no.2
/
pp.170-183
/
2017
Bacterial fruit blotch (BFB), which is caused by Acidovorax citrulli, is a serious threat to watermelon growers around the world. The present study was conducted to screen effective rhizobacterial isolates against 35 different A. citrulli isolates and determine their efficacy on BFB and growth parameters of watermelon. Two rhizobacterial isolates viz. Paenibacillus polymyxa (SN-22), Sinomonas atrocyanea (NSB-27) showed high inhibitory activity in the preliminary screening and were further evaluated for their effect on BFB and growth parameters of three different watermelon varieties under greenhouse conditions. The greenhouse experiment result revealed that SN-22 and NSB-27 significantly reduced BFB and had significant stimulatory effect on total chlorophyll content, plant height, total fresh weight and total dry weight compared to uninoculated plants across the tested three watermelon varieties. Analysis of the 16S ribosomal RNA (rRNA) sequences revealed that strains SN-22 belong to P. polymyxa and NSB-27 to S. atrocyanea with the bootstrap value of 99% and 98%, respectively. The isolates SN-22 and NSB-27 were tested for antagonistic and PGP traits. The result showed that the tested isolates produced siderophore, hydrolytic enzymes (protease and cellulose), chitinase, starch hydrolytic enzymes and they showed phosphate as well as zinc solubilizing capacity. This is the first report of P. polymyxa (SN-22) and S. atrocyanea (NSB-27) as biocontrol-plant growth promoting rhizobacteria on watermelon.
We investigated the effect of infection time of CGMMV on the growth and quality of watermelon and cucumber plants. The effect (damages by CGMMV) was estimated on the watermelon where CGMMV had been inoculated at different growth stages, vegetative (transplanting stage, vegetative growth stage) and reproductive growth stage (fruiting stage and fruit hypertrophy stage). In the case of cucumber, CGMMV was inoculated at transplanting stage and Erst flowering stage, respectively. When watermelon was infected with CGMMV at vegetative growth stage, vine length, internode length, leaf area, and fruit weight of the plants largely decreased compared with control plants, while the infected plant growth was not very different from control plants when it was infected at reproductive growth stage. Brix of the fruit of watermelon also decreased when the plants was infected with the virus earlier than fruiting stage. The occurrence of 'Pisubag', internal discoloration and decomposition of watermelon fruits, tended to be increased as earlier infection time with CGMMV In the case of cucumber infection time with CGMMV did not influence earlier growth of the plants, but did later growth showing that plant height, vine length, internode length, number of leaf, leaf wide, and leaf length of the plants decreased as infection time became to be earlier.
The selected five plant growth-promoting rhizobacteria (PGPR) strains, WR8-3 (Pseudomonas fluorescens), WR8-6 (P. putida), WR9-9 (P. fluorescens), WR9-11 (Pseudomonas sp.), and WR9-16 (P. putida) isolated in the rhizosphere of watermelon plants were tested on their growth promotion and control effect against gummy stem rot of watermelon. Strains, WR8-3 and WR9-16 significantly increased stem length of watermelon, and there was a little increase in leaf area, fresh weight and root length when strains, WR8-3, WR9-9 and WR9-16 were treated. Generally, seed treatment was better for plant growth promotion than the soil drench, but there was no significant difference. Seed treatment and soil drench of each bacterial strain also significantly reduced the mean lesion area (MLA) by gummy stem rot, but there was no significant difference between the two treatments. At initial inoculum densities of each strain ranging from 10$^6\;to\;10^{15}$ cfu/g seed, approximately the same level of disease resistance was induced. But resistance induction was not induced at the initial inoculum density of 10$^3$ cfu/g seed. Resistance was induced by treating the strains, WR9-9, WR9-11 and WR9-16, on all of four watermelon varieties tested, and there was no significant difference in the decrease of gummy stem rot among varieties. Populations of the strains treated initially at log 9-10 cfu/g seed, followed with a rapid decrease from planting day to 1 week after planting, but the population density was maintained above log 5.0 cfu/g soil until 4 weeks after planting. Generally no or very weak in vitro antagonism was observed at the strains treated excepting WR9-11. Rifampicin-resistant bacteria which had been inoculated were not detected in the stems or leaves, which suggesting that the bacterium and the pathogens remained spatially separated during the experiment. This is the first report of rsistance induction in watermelon to gummy stem rot by PGPR strains.
Phylogenetic relationships in Korean watermelons were evaluated by genetic similarity coefficients using 15 SSR(simple sequence repeat), 14 SCAR(sequence characterized amplified region) and 14 CAPS(sequence characterized amplified region) markers. The SSR markers were selected from previously reported melon and watermelon SSRs through testing polymorphisms within a set of commercial $F_1$ varieties. The SCAR and CAPS markers were developed from polymorphic AFLP(amplified fragment length polymorphism) markers between inbred lines 'BN4001' and 'BN4002'. From the AFLP analysis, 105 polymorphic fragments were identified between the inbred lines using 1,440 primer combinations of EcoRI+CNNN and XbaI+ANNN. Based on the sequencing data of these polymorphic fragments, we synthesized sequence specific primer pairs and detected clear and reliable polymorphisms in 27 primer pairs by indels(insertion/deletion) or RFLP(restriction fragment length polymorphism). A total of 43 sequence-based PCR markers were obtained and polymorphic information content(PIC) was analyzed to measure the informativeness of each marker in watermelon varieties. The average PIC value of SCAR markers was 0.41, which was similar to that of SSR markers. Genetic diversity was also estimated by using these markers to assess the phylogenetic relationships among commercial varieties of watermelon. These markers differentiated 26 Korean watermelon varieties into two major phylogenetic groups, but this grouping was not significantly correlated with their morphological and physiological characteristics. The mean genetic similarity was 66% within the complete set of 26 commercial varieties. In addition, these sequence-based PCR markers were reliable and useful to identify cultivars and genotypes of watermelon.
Melon necrotic spot virus (MNSV) was recently identified on watermelon (Citrullus vulgaris) in Korea, displaying as large necrotic spots and vein necrosis on the leaves and stems. The average occurrence of MNSV on watermelon was found to be 30-65% in Hapcheon and Andong City, respectively. Four isolates of the virus (MNSV-HW, MNSV-AW, MNSV-YW, and MNSV-SW) obtained from watermelon plants in different areas were non-pathogenic on ten general indicator plants, including Chenopodium quinoa, while they infected systemically six varieties of Cucurbitaceae. The virus particles purified by 10-40% sucrose density gradient centrifugation had a typical ultraviolet spectrum, with a minimum at 245 nm and a maximum at 260 nm. The morphology of the virus was spherical with a diameter of 28-30 nm. Virus particles were observed scattered throughout the cytoplasm of watermelon cells, but no crystals were detected. An ELISA was conducted using antiserum against MNSV-HW; the optimum concentrations of IgG and conjugated IgG for the assay were $1{\mu}l/ml$ and a 1:8,000-1:10,000 dilutions, respectively. Antiserum against MNSV-HW could capture specifically both MNSV-MN from melon and MNSV-HW from watermelon by IC/RT-PCR, and they were effectively detected with the same specific primer to produce product of 1,172 bp. The dsRNA of MNSV-HW had the same profile (4.5, 1.8, and 1.6 kb) as that of MNSV-MN from melon. The nucleotide sequence of the coat protein of MNSV-HW gave a different phylogenetic tree, having 17.2% difference in nucleotide sequence compared with MNSV isolates from melon.
The purpose of this study was to evaluate cookies prepared with five different quantities (0, 5, 10, 15, and 20%) of watermelon powder (WP). This study analyzed quality characteristics, consumer liking, and CATA (check-all-that-apply) of the samples. The density and pH of the cookie dough and the L-value of the cookies tended to decrease as the amount of watermelon powder increased (p<0.05), whereas the spread factor, a-value, and hardness of the cookies tended to increase as the amount of watermelon powder increased (p<0.001). The b-value tended to increase up to WP10, but it tended to decrease from WP15 (p<0.001). The results of the evaluation of consumer liking showed that overall liking was the highest for WP5 and lowest for WP20 (p<0.05). In the analysis of the CATA survey, the main reasons for liking for all the samples were 'Appearance', 'Color', 'Sweet taste', 'Nutty odor/flavor', 'Crispiness', and 'Familiarity'. WP5 showed the most diverse reasons for being liked. The common reason for disliking samples with the addition of watermelon powder was 'Residual feel in the mouth'. In the correspondence analysis, attributes of 'Stale taste', 'Clean taste', 'Softness', 'Familiarity', 'Moistness', 'Color', 'Blandness' were detected in the WP0 and WP5. The results indicate that WP5 with a 5% supplementation of watermelon powder is appropriate for improving the quality and consumer acceptability of the cookies.
Selection of yeast strains, optimum conditions for alcohol fermentation, sterilization methods, and additives for improving wine quality were investigated to manufacture watermelon wine. Eight yeast strains exhibited significant alcohol fermentation, among which KWS 06 was selected for watermelon wine fermentation, because watermelon wine made by this strain showed the best overall acceptability in sensory evaluation. Sucrose was determined as the best saccharide for alcohol fermentation among sucrose, corn syrup, glucose, fructose, and lactose. Optimum concentration of soluble solid and $(NH_{4})_{2}HPO_{4}$ of nitrogen source were $24^{\circ}Brix and 0.2%, respectively. Addition of raspberries and omija increased wine flavor and alcohol production, respectively, with optimum alcohol production, taste, and color achieved with addition of 20 g/L raspberries and 10 g/L omija. Best sensory quality was obtained by addition of 0.04 % watermelon flavorant to the juice.
We investigated the occurrence pattern of diseases and insect pests and disease control efficacy of organic materials against watermelon powdery mildew in small type watermelon in Eumseong, Chungcheongbuk-do, 2015. The result of this study, the small type watermelon was damaged by diseases and pests such as Didymella bryoniae, Podosphaera xanthii, Aphis gossypii, Tetranychus urticae, thrips, Spodoptera exigua and Spodoptera litura. Among them, the occurrence of P. xanthii, T. urticae and thrips was high. Diseased leaf rate by P. xanthii, a casual agent of powdery mildew, was 27~99.3% in three small type watermelon cultivars in the middle of June. The number of T. urticae per leaf was high from 79.9 to 111 in three small type watermelon cultivars in the middle of June. It showed high number of thrips captured by yellow and blue sticky trap. Highest numbers of yellow sticky trap (407) and of blue sticky trap (774) were detected in the middle and first of June, respectively. The disease control efficacy of mayonnaise, oleic acid and three organic materials against powdery mildew of small type watermelon in fields was evaluated. As thre results, the symptoms of plant disease were effectively reduced by over 60% in the treatments of materials such as sodium bicarbonate 80%, mayonnaise and the extract of Rheum palmatum 1%. The highest control efficacy was 83% in the treatment of sodium bicarbonate 80%. From this study, we had a information of the occurrence pattern of diseases and insect pests in small type watermelon and the treatment of material containing sodium bicarbonate 80% was very effective for controlling against powdery mildew.
Watermelon production is often limited by powdery mildew in areas with a large daily temperature range. Development of resistant watermelon cultivars can protect against powdery mildew; however, little is known about the characteristics of its causal agents. Here, we identified the genus and race of a causal pathogen of powdery mildew in Ansung province of South Korea, and developed molecular markers for the generation of resistant watermelon cultivars. The causal pathogen was determined to be Podosphaera xanthii based on multiple sequence alignments of internal transcribed spacers (ITS) of rDNA. The physiological race was identified as 1W, and the Ansung isolate was named P. xanthii 1W-AN. Following inoculation with the identified P. xanthii 1W-AN, we found inheritance of the resistant gene fitting a single dominant Mendelian model in a segregated population ('SBA' ${\times}$ PI 254744). To develop molecular markers linked to fungus-resistant loci, random amplified polymorphic DNA (RAPD) was accomplished between DNA pooled from eight near-isogenic lines (NILs; $BC_4F_6$), originated from PI 254744 and susceptible 'SBB' watermelon. After sequencing bands from RAPD were identified in all eight NILs and PI254744, 42 sequence-characterized amplifiedregion (SCAR) markers were developed. Overall, 107 $F_2$ plants derived from $BC_4F_6$ NIL-1 ${\times}$ 'SBB' were tested, and one SCAR marker was selected. Sequence comparison between the SCAR marker and the reference watermelon genome identified three Nco I restriction enzyme sites harboring a single nucleotide polymorphism, and codominant cleavage-amplified polymorphic site markers were subsequently developed. A CAPS marker was converted to a high-resolution melt (HRM) marker, which can discriminate C/T SNP (254PMR-HRM3). The 254PMR-HRM3 marker was evaluated in 138 $F_{2:3}$ plants of a segregating population ('SBA' ${\times}$ PI254744) and was presumed to be 4.3 cM from the resistance locus. These results could ensure P. xanthii 1W-AN resistance in watermelon germplasm and aid watermelon cultivar development in marker-assist breeding programs.
Dilution concentration of watermelon juice, concentrations of added sugar, citric acid, and vitamin C, sterilization temperature and time, and natural color extracts were evaluated to determine optimum conditions for watermelon beverage production. Optimum dilution concentration of watermelon juice and optimum content of soluble solid were 40% and $12^{\circ}Brix$, respectively. Addition of 0.5 and 0.3 g/L or 1.0 and 0.3 g/L citric acid and vitamin C gave optimum sensory quality. Sterilization of watermelon beverage at above $70^{\circ}C$ decreased redness. Sterilization at $60^{\circ}C$ for 10 to 30 min or at $70^{\circ}C$ for 10 min achieved best sensory quality. Addition of 20 g/L raspberries gave best sensory quality among raspberries, omija, and borage. Hot water was better than alcohol for extraction of natural color. Ratio of extracts for optimum sensory quality was 7 : 3 for extract of 20 g raspberries/L : extract of 30 g omija/L.
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