Detection of gene flow from GM to non-GM watermelon in a field trial

Gene flow from genetically modified (GM) crops to conventional non-GM crops is a serious concern for protection of conventional and organic farming. Gene flow from GM watermelon developed for rootstock use, containing cucumber green mottle mosaic virus (CGMMV)-coat protein (CP) gene, to a non-GM isogenic control variety “Clhalteok” and grafted watermelon “Keumcheon” was investigated in a small scale field trial as a pilot study. Hybrids between GM and non-GM watermelons were screened from 1304 “Chalteok” seeds and 856 “Keumcheon” seeds using the duplex PCR method targeting theCGMMV- CP gene as a marker. Hybrids were found in all pollen recipient plots. The gene flow frequencies were greater for “Chaiteok” than for “KeumcheonD; with 75% outcrossing in the “Chaiteok” plot at the closest distance (0.8 m) to the GM plot. A much larger scale field trial is necessary to identify the isolation distance between GM and non-GM watermelon, as the behaviors of insect pollinators needs to be clarified in Korea.     

Effects of photo-oxidative loss of FeNa2EDTA and of higher iron supply on chlorophyll content, growth and propagation rate in triploid watermelon cultures

Summary Triploid watermelon (Citrullus lanatus [Thunb.] Matsum. & Nakai) Arka Manik cultures exhibited a general pale color and yellowing of lower leaves by 3–4 wk following subculturing to fresh medium. Protecting the medium from light by covering the surface with sterile aluminum foil significantly improved leaf color and chlorophyll content by 50–60%. Raising the concentration of FeNa₂EDTA (Fe-EDTA) in the light-exposed medium from the normal level (100 μM) to 150 or 200 μM significantly increased the leaf chlorophyll two- to threefold, suggesting that the light-induced effect was mainly attributable to photo-oxidation of Fe-EDTA. Fe-EDTA supplementation also resulted in better expansion and lobe formation of upper leaves, a reduction in internode length, an increase in the number of nodes, and a significant improvement in propagation rate. There was also a reduction in axillary sprout growth at elevated Fe-EDTA levels, indicating a more prominent apical dominance effect and a reduction in the number and vigor of tendrils. Increasing the Fe-EDTA level in the medium to 150 μM is suggested for routine micropropagation of triploid watermelon, in order to compensate for the photo-oxidative loss of Fe-EDTA.     

Somatic embryogenesis and plant regeneration from immature cotyledons of watermelon

Cotyledon expiants from immature embryos of five watermelon [Citrullus lanatus (Thunb.)Matsum. & Nakai] genotypes were incubated in the dark for three weeks on a modified MS medium containing B5 vitamins, 2,4-D (10, 20 or 40M), 0.5 M of either BA or TDZ, and 7 g·1^-1 TC agar. Somatic embryos, some with well developed cotyledons, were observed on cotyledon expiants three to four weeks after transfer to MS medium without PGRs and 16h photoperiod. The best PGR combination for somatic embryogenesis was 10 M 2,4-D and 0.5 M TDZ Somatic embryogenesis was greatest (30%) when cotyledon expiants were established from 18-day-old immature embryos. Somatic embryos were germinated on MS medium without PGRs. Plants were transferred to Magenta boxes containing ProMix for three weeks before being transplanted to the field where they formed fertile male and female flowers that produced normal fruit.Abbreviations PGR plant growth regulator - BA benzyladenine - TDZ thidiazuron - 2,4-D 2,4-dichlorophenoxyaceticacid - NAA -naphthaleneacetic acid - 2,4,5-T 2,4,5-trichlorophenoxyacetic acid     

Changes in soluble carbohydrates during phytochrome-regulated petiole elongation in watermelon seedlings

Changes in soluble carbohydrate composition and concentration in leavesand petioles of watermelon (Citrullus lanatus (Thunb)Matsum and Nakai cv. Sugar Baby) seedlings during early stages ofphytochrome-regulated petiole elongation were investigated. Watermelon seedlingswere grown in a controlled environment with 350 molm^–2 s^–1 photosynthetically activeradiation (PAR) during a 12-h photoperiod. Low intensity end-of-day(EOD) light treatments (for 15 min) of red (R), far-red (FR) and FRfollowed by R (FR/R) were initiated when the seedlings were 14 days old.Seedling growth, and soluble carbohydrate concentration and composition inleaves and petioles were determined after 3 and 6 days of EOD light treatments.The EOD FR increased the petiole length and dry mass partitioned to petioles asearly as 3 days into the treatment. This increased petiole dry mass inFR-treated plants was accompanied with an increase in reducing sugar (glucoseand fructose) concentration in the petioles. Although both leaves and petiolesshowed this effect, the relative increase was greater in petioles than leaves.While the most abundant sugars in petioles were fructose and glucose, thepredominant sugars in leaves were sucrose, raffinose, and stachyose. Thephotoreversion of FR induced changes in growth and sugar concentrations by Rindicates the involvement of phytochrome in these processes.     

Dark pretreatment improves adventitious shoot organogenesis from cotyledons of diploid watermelon

The influence of light incubation during embryo germination on shoot organogenesis from cotyledons of four diploid watermelon [Citrullus lanatus (Thumb.) Matsum. & Nakai cultivars was examined. Germinating embryos in darkness significantly improved the number of explants that produced harvestable shoots during the 6 week incubation period on shoot regeneration medium under a 16-h photoperiod. The percentage of explants with shoots more than doubled for `Crimson Sweet' and was about 1.5-fold greater for `Sweet Gem' and `Yellow Doll' when embryos were germinated in darkness. The percentage of explants with shoots was not significantly improved for `Minilee' by pretreating seedlings in darkness. This study demonstrates that optimal shoot regeneration can be obtained by germinating embryos in darkness before preparing cotyledon explants. This revised version was published online in June 2006 with corrections to the Cover Date.     

High-frequency shoot regeneration from cotyledon explants of watermelon cv. sugar baby

Summary A protocol for in vitro shoot regeneration from cotyledon explants of Citrullus lanatus (Thunb.) Matsum. & Nakai cv. Sugar Baby is described. The cotyledons excised from 7-d-old aseptic seedlings showed the highest percentage of shoots on Murashige and Skoog (MS) + N⁶-benzyladenine (BA; 3.0 μM) + N⁶-[2-isopentenyl] adenine (2iP; 3.0 μM) and MS + BA (3.0 μM) + indole-3-acetic acid (IAA; 3.0 μM). Whereas the latter medium induced shoot regeneration after the callusing of the explant, the former stimulated direct shoot formation. The regenerated shoots were rooted and the resulting plants were established in earthen pots with 55% success.     

Nutrient utilization in liquid/membrane system for watermelon micropropagation

Watermelon [Citrullus lanatus (Thunberg) Matsumura and Nakai] proliferating shoot meristems from established shoot cultures were inoculated on modified Murashige and Skoog salts medium supplemented with 10 μM 6-benzyladenine (BA) for shoot proliferation and on similar medium supplemented with 1 μM BA and 10 μM gibberellic acid (GA₃) for shoot elongation. Agar-solidified medium and microporous polypropylene membrane rafts in liquid medium were used to support the tissues. Growth over culture time of proliferating and elongating tissues in liquid and agar-solidified media were compared. Nutrient depletion in liquid medium was monitored and quantified using ion selective electrodes. Tissue fresh weights in both proliferation and shoot elongation media were greater in liquid than in agar-solidified medium. Relative dry matter content, however, was greater in agar-solidified than in liquid medium. More shoots elongated in agar-solidified than in liquid medium. The numbers of buds or unelongated shoot meristems, however, were comparable for both the liquid and agar-solidified medium. Proliferating and elongating tissues in liquid medium used Ca⁺⁺ and K⁺ minimally. NO ₃ ⁻ was utilized but not depleted by proliferating tissues. NH ₄ ⁺ , however, was depleted. Most of the NH ₄ ⁺ was utilized by the proliferating tissues within 21 days of culture when growth rate was greatest. At 35 days, residual Ca⁺⁺, K⁺, NO ₃ ⁻ , and NH ₄ ⁺ in proliferation medium were 81.0%, 67.8%, 55.7%, and 1.2% of initial levels, respectively. NO ₃ ⁻ and NH ₄ ⁺ in shoot elongation medium were depleted. The greatest NO ₃ ⁻ and NH ₄ ⁺ utilization was observed during the first 14 days of culture when the largest growth rate was obtained. The residual Ca⁺⁺, K⁺, NO ₃ ⁻ , and NH ₄ ⁺ in shoot elongation medium at 38 days were 63.5%, 37.9%, 21.2%, and 24.3% of initial concentrations, respectively. At the end of experiment, 72.3% and 42.8% of initial sugars were still remaining in the shoot proliferation and shoot elongation medium, respectively. Technical Contribution No. 3236 of the South Carolina Agricultural Experiment Station.     

Effect of the synthetic auxin 2,4-DP on fruit development of loquat

Application of the butylglycol ester of 2,4-DP increased final fruit size in loquat without causing fruit thinning. Its effectiveness depended on the concentration applied and treatment date. When applied at the onset of the cell enlargement fruit stage, 25 mg l^–1 was the most effective treatment. Fruit diameter distribution showed a significant shift to the larger size for treated trees; further, fruit colour break and maturation were encouraged and harvest time was earlier than in untreated trees.     

Effect of pollination intensity on fruit and seed set in cacao (Theobroma cacao L.)

We studied the functional relationship between pollination intensity and fruit survival as well as the number of seeds per pod in the tropical tree Theobroma cacao L. on a Forastero Upper-Amazon clone (UPA 409) in Ivory Coast. Cutting the style 24 h after pollination allowed for counting the number of pollen grains deposited on a stigma without affecting fruit set and seed development. Forty-three pollen grains were necessary to reach 50% of maximum fruit set 28 days after pollination. Above 115 pollen grains, the proportion of developing ovaries reached a maximum of 88% 28 days after pollination and 75% at maturity. With fewer than 238 pollen grains per stigma, there was a close relationship between pollination intensity and number of seeds per pod; the pollenseed ratio increased from 1.61 to 3.81 for PI increasing from 30 to 238 pollen grains. For higher pollination intensities, the average number of seeds per pod reached a maximum of 58. The relationship between pollination intensity and seed content was modelled. Results are consistent with the hypothesis that ovules attracted pollen tubes in a similar way regardless of whether or not they had already been reached by another pollen tube.     

Ribulose-1,5-bisphosphate carboxylase and fruit set or degeneration of unpollinated ovaries of Pisum sativum L.

The polypeptide patterns obtained by sodium dodecylsulphate-polyacrylamide gel electrophoresis of undigested and autodigested extracts from pea (Pisum sativum L.) ovaries at the early stages of development or degeneration have been studied. Development of unpollinated ovaries was stimulated by application of different plant growth regulators (gibberellic acid, 2,4-dichlorophenoxyacetic acid, and N⁶-benzyladenine) or by plant topping. Polypeptide bands of similar mobility to ribulose-1,5-bisphosphate carboxylase (RuBPCase) subunits (16 and 55 kDa) could be detected in all types of autodigested extracts from stimulated ovaries. However these bands were absent in electrophoretic patterns of autodigested extracts from unstimulated ovaries after 3 d post anthesis and in patterns of autodigested mixtures of these extracts with either those from stimulated ovaries or those from unstimulated ovaries before day 3. These observations indicate that a proteolytic activity which promotes the hydrolysis of RuBPCase appears in unstimulated ovaries about 3 d after anthesis. This event coincides with the loss of the capacity of unpollinated ovaries to develop in response to gibberellic acid and with the degeneration of the ovary wall.Abbreviations BA N⁶-benzyladenine - 2,4-D 2,4-dichlorophenoxyacetic acid - GA₃ gibberellic acid - RuBPCase ribulose-1,5-bisphosphate carboxylase - SDS-PAGE sodium dodecylsulphate-polyacrylamide gel electrophoresis     

An archaeobotanical contribution to the history of watermelon,<Emphasis Type="Italic"> Citrullus lanatus</Emphasis> (Thunb.) Matsum. &; Nakai (syn.<Emphasis Type="Italic"> C. vulgaris</Emphasis> Schrad.)

The discovery of several 5000-year old seeds of wild watermelon, Citrullus lanatus, at an archaeological site Uan Muhuggiag in southwest Libya, re-opens the debate on the origin, wild distribution and domestication history of this species. The seeds were found within a plant assemblage of wild seeds and fruits, associated with pottery and bones of domestic animals belonging to Neolithic pastoralists. The presumed wild progenitor of the modern cultivar C. lanatus is today found exclusively in a region centring on the Kalahari Desert. This new archaeobotanical record raises the possibility that this distribution was much more extensive in the past.     

Between-year variation in flowering and fruit set in frost-prone and frost-sheltered populations of dioecious Rubus chamaemorus

Summary The flowering and fruiting patterns of the dioecious perennial herb Rubus chamaemorus L. were studied in frost-prone (open) and frost-sheltered (Shaded) habitats in northern Sweden over 6 years. The number of ramets with flower buds, the proportion of flower buds that opened, and fruit set varied markedly between years. In the frost-prone populations, the occurrence or absence of detrimental frosts during the development of flowers and fruit could explain much of the variation, both in the proportion of flower buds that developed into flowers, and in fruit set. In the frost-sheltered populations, most female flowers that did not develop into fruit aborted without any signs of physical damage and before any ovules had begun to enlarge. Flower mortality caused by herbivores feeding on reproductive parts was commonly low, but reached values higher than 10% in one of the shaded populations. Hand-pollination increased the proportion of ovules producing seeds in the mature fruits by about 20%, and in one year also increased fruit set significantly in one population. Fruit-producing female ramets had a higher mortality and a lower probability of flowering in the subsequent year than male ramets and non-fruiting female ramets. In R. chamaemorus, the conditions for fruit maturation are highly unpredictable at the time of flower initiation. It is suggested that the apparent over-initiation of flower buds is advantageous, as it allows the plant to attain a high reproductive success in years favourable for flowering and fruit development.     

Single nucleotide polymorphisms generated by genotyping by sequencing to characterize genome-wide diversity,linkage disequilibrium,and selective sweeps in cultivated watermelon

Background

A large single nucleotide polymorphism (SNP) dataset was used to analyze genome-wide diversity in a diverse collection of watermelon cultivars representing globally cultivated, watermelon genetic diversity. The marker density required for conducting successful association mapping depends on the extent of linkage disequilibrium (LD) within a population. Use of genotyping by sequencing reveals large numbers of SNPs that in turn generate opportunities in genome-wide association mapping and marker-assisted selection, even in crops such as watermelon for which few genomic resources are available. In this paper, we used genome-wide genetic diversity to study LD, selective sweeps, and pairwise F_ST distributions among worldwide cultivated watermelons to track signals of domestication.

Results

We examined 183 Citrullus lanatus var. lanatus accessions representing domesticated watermelon and generated a set of 11,485 SNP markers using genotyping by sequencing. With a diverse panel of worldwide cultivated watermelons, we identified a set of 5,254 SNPs with a minor allele frequency of ≥ 0.05, distributed across the genome. All ancestries were traced to Africa and an admixture of various ancestries constituted secondary gene pools across various continents. A sliding window analysis using pairwise F_ST values was used to resolve selective sweeps. We identified strong selection on chromosomes 3 and 9 that might have contributed to the domestication process. Pairwise analysis of adjacent SNPs within a chromosome as well as within a haplotype allowed us to estimate genome-wide LD decay. LD was also detected within individual genes on various chromosomes. Principal component and ancestry analyses were used to account for population structure in a genome-wide association study. We further mapped important genes for soluble solid content using a mixed linear model.

Conclusions

Information concerning the SNP resources, population structure, and LD developed in this study will help in identifying agronomically important candidate genes from the genomic regions underlying selection and for mapping quantitative trait loci using a genome-wide association study in sweet watermelon.

Electronic supplementary material

The online version of this article (doi:10.1186/1471-2164-15-767) contains supplementary material, which is available to authorized users.     

位置效应对巫山淫羊藿花蜜分泌和结实的影响

位置效应在开花植物中广泛存在。以野生巫山淫羊藿种群为研究对象,运用单因素方差分析法,研究位置效应对结实性和花蜜分泌的影响。结果表明:三个巫山淫羊藿种群不同位置的结实率显著差异,其中基部的结实率明显大于中部和顶部; 位置效应对果实重量及果实长、宽的影响均显著,表现出基部>中部>顶部的趋势; 总状花序上不同开花位置对单个果实种子数的影响显著,且基部大于顶部。但位置效应对单个果实种子败育率的影响不显著,均值呈现顶部>中部>基部变化规律; 巫山淫羊藿不同开花位置的花蜜常备量和含糖量的差异显著。三个种群基部的花蜜常备量较高,而花蜜含糖量在顶部较高。因此,对巫山淫羊藿野生种群进行研究与保护时应注意位置效应对生殖成功的影响,选择基部果实进行试验或育种,应维持种群多样性及稳定性。     

不同生境与花部特征对巫山淫羊藿结实特性的影响

对不同生境中巫山淫羊藿结实特性进行比较研究。结果表明:(1)三个种群在繁殖时期中不同花部特征的植株繁殖投资差异不大,如每株分枝数、每株花序数和每株开花数差异均不显著;(2)三个种群中每株结实数、每株结实率、每花序结实数在不同花部特征中差异显著,PLS和SAA要显著大于PESS;(3)三种花部特征在不同生境中单株分枝数和单株花序数均无显著差异。在不同花部特征中种群1开花数量相较少,因而在结实方面均值均要小于种群2和种群3;(4)不同花部特征的果实饱满种子数、败育数和败育率均具有显著差异,而种子数差异不大,但在种群间均无显著的差异。说明种子的多少更多地受到花部特征的影响。总之,巫山淫羊藿的结实特性受到生境和花部特征的双重影响,其中花型比生境对巫山淫羊藿结实特性的影响更为明显。     

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.     

Effect of ovular receptivity on seed set and fruit development in cucumber (Cucumis sativus L.)

Summary Seed set and fruit development in cucumber (Cucumis sativus L.) were studied in relation to female flower receptivity from day — 2 before anthesis to day + 2 after anthesis. The female cucumber flower is protogynous. The pistil was receptive 2 days before anthesis. The iso-electric focusing (IEF) patterns of the stigma/style proteins, were identical from day -5 to day +2. In pollinated flowers in vivo germination and pollen-tube growth in the ovary were affected by pistil age from day -2 to day +2. In addition, differences in sectorial filling in full seeds were observed within the fruits. A negative correlation was observed between the frequency of fertilized ovules in the pedoncular part of the fruit and ovary length at the time of pollination. In the whole fruit, significant differences in the number of full seeds and fruit size at maturity were found, and these were observed to be correlated with the various stages of female flower maturation at pollination. The day -2 and day +2 stages yielded the smallest fruits with few full seeds compared to the day -1, day 0 and day +1 stages, which had the biggest fruits and a large number of full seeds. A strong positive correlation was found between total seed number (including full and empty seeds), fruit length and weight at maturity. All these results suggest that both seed set in the different parts of the fruit and fruit development are controlled by ovular receptivity rather than by stigma/style receptivity.     

镉胁迫对西瓜幼苗生长及其叶片解剖结构和生理特性的影响

该研究以‘北抗1号’西瓜幼苗为试验材料,采用实验室盆栽方法,考察在不同浓度镉胁迫(0、60、120、180和240 mg/L)处理下西瓜幼苗的生长及其叶片生理特性和解剖结构的变化特征,初步探讨西瓜耐受镉胁迫的生理机制。结果显示：(1)在镉胁迫条件下,西瓜幼苗的生长受到抑制,随着镉胁迫浓度的增加,西瓜幼苗的叶片形态黄化现象逐渐加重,根系形态逐渐纤弱,株高、茎粗、茎节数和叶片数均呈现下降趋势。(2)镉胁迫下,西瓜幼苗叶片主脉中细胞受损,主脉直径显著减小,叶肉组织疏密度显著降低。(3)随着镉胁迫浓度的升高,西瓜幼苗的含水量、净光合速率、SOD活性等显著降低,丙二醛和游离脯氨酸含量显著升高,POD活性和可溶性蛋白含量先升高后降低,生理特性受到显著影响。研究发现,‘北抗1号’西瓜幼苗对镉有一定的适应性,在低浓度(60 mg/L)镉胁迫处理下,西瓜幼苗的形态特征、生理特性变化不显著,但在高浓度(180 mg/L)镉胁迫下,幼苗的生长受到严重抑制,渗透调节系统以及生物膜保护系统严重受损。     

Identification of the double genome donor in spontaneous triploid tomato plants by RFLP analysis

Independent spontaneous triploid tomato plants (Lycopersicon esculentum Mill.) were collected among diploid hybrids growing in commercial greenhouses. Ploidy levels were verified by counting chromosomes, and the donor of the double genome dose was determined by restriction fragment length polymorphism (RFLP) analysis. The TG101 probe, which is tightly linked to the Tm-2 ^a locus, revealed different restriction patterns between TMV-resistant and TMV-susceptible parent lines. The parent donor which provided two genomes to the triploid was identified by comparing the relative intensity of alleles in the triploid with that in the diploid. The results indicate that both parents can serve as a double genome donor.     

Factors affecting fruit set in Aizoaceae species of the Succulent Karoo

By reducing the number of flowers and fruits, intense grazing activities of domestic livestock are considered to have negative effects on the plant recruitment of perennial plants in the Succulent Karoo, South Africa. In the present study, the fruit set of six perennial species belonging to the Aizoaceae was investigated under two different grazing intensities. Two species (Cheiridopsis imitans and Leipoldtia schultzei) were examined more closely to determine whether factors other than grazing impact fruit production of these plants. Apart from reproductive output, the population structure of these two species was explored in heavily and slightly grazed areas.For all investigated species, heavy livestock grazing was identified as a major threat to fruit production. Deviations from this pattern were recorded in a drought year, when harsh weather conditions reduced herd sizes dramatically and the usually high grazing pressure was alleviated. However, detailed analyses revealed that also other biotic factors can cause severe fruit and flower losses even exceeding that caused by grazing. Caterpillars of the noctuid moth, Diaphone eumela, and two rodent species, Otomys unisulcatus and Rhabdomys pumilio, proved to be further important herbivores of flowers and fruits. Experiments with caged plants, excluding livestock from grazing, confirmed though that livestock grazing definitely accounts for a continuous reduction of fruit production. Insufficient pollination success was another parameter reducing fruit set; however, this was almost exclusively observed under low grazing pressure.