Effects of Forchlorfenuron on the Morphology, Metabolite Accumulation, and Transcriptional Responses of Siraitia grosvenorii Fruit.

Siraitia grosvenorii fruit, called luo-han-guo (LHG), have been used as a traditional Chinese medicine (TCM) and dietary supplements for many years. Mogrosides, the main bioactive ingredients in LHG, are commercially available worldwide as a non-sugar-based and noncaloric sweetener. However, the production cannot meet the increasing market demand because of the low content of mogrosides and the small size of LHG. Therefore, some advanced technologies have been applied for improving the quality of LHG. Forchlorfenuron (CPPU), a plant growth regulator, is widely applied to promote plant yield and the secondary metabolite synthesis. Here, the content of nine mogrosides and three intermediates in LHG that were treated with three different concentrations of CPPU were determined by LC-MS/MS and GC-MS, respectively. The total content of mogrosides in LHG treated with CPPU was not enhanced, and the proportion of some main bioactive ingredients, including mogroside V (MV), were decreased relative to that of the control treatment. Morphological and cytological observations showed CPPU could make an early lignification in fruit epidermal cells, and 5 or 25 mg L-1 CPPU could inhibit LHG growth. The expression levels of 24 key genes in the mogroside biosynthesis pathway were measured and revealed that genes downregulated in upstream, and different expressions of SgUGTs would affect the accumulations and proportions of mogrosides in LHG induced by CPPU. This was the first study that applied CPPU individually on LHG, and assessed effects of CPPU on the morphology, the accumulation of metabolites, and expression profiles of 24 structural genes. The CPPU effects on LHG were undesirable, including development inhibition and the decrease of main mogroside content. These will provide guidance for the rational application of CPPU.

Calcium chloride and 1-methylcyclopropene treatments delay postharvest and reduce decay of New Queen melon.

In this study, newly harvested New Queen melons were treated with calcium chloride (CaCl2) and 1-methylcyclopropene (1-MCP) alone or in combination before storage. The results showed that the respiration rate, ethylene release, the activity and gene expression of pectinases such as polygalacturonase (PG), pectin methylesterase (PME) and pectate lyase (PL) in New Queen melons were dramatically decreased by treatments with 0.18 mol/L CaCl2 and/or 1 µL/L 1-MCP. Meanwhile, the climacteric behavior and flesh hardness reduction were inhibited. We also found that softer melon flesh was more conducive to the growth and reproduction of decay-causing microorganisms according to their growth curves in melons that were different in flesh hardness, suggesting inhibiting fruit softening can slow down the growth of microorganisms in fruit flesh, and thus reduce fruit decay rate. The combined use of CaCl2 and 1-MCP was more effective in suppressing respiration rate, ethylene release and protopectin hydrolysis, which could greatly delay the softening, reduce the decay rate, and extend the shelf life of New Queen melons.

Thermal Stress in Melon Plants: Phytoprostanes and Phytofurans as Oxidative Stress Biomarkers and the Effect of Antioxidant Supplementation.

The extreme temperatures generated in the melon crop, early harvest, induce an increase in reactive oxygen species (ROS) plant levels leading to oxidative stress. Phytoprostanes (PhytoPs) and phytofurans (PhytoFs) are plant metabolites derived from α-linolenic acid oxidation induced by ROS. The aims of this work were to evaluate PhytoPs and PhytoFs as oxidative stress biomarkers in leaves of melon plants thermally stressed. In addition, to fortify melon plant antioxidant defenses, foliar spraying was assayed using salicylic and gallic acid solutions and Ilex paraguariensis extract. PhytoP and PhytoF concentration ranges were 109-1146 and 130-4400 ng/g, respectively. Their levels in stressed plants were significantly higher than in nonstressed samples. In stressed samples treated with I. paraguariensis, PhytoP and PhytoF levels were significantly lower than in stressed samples without antioxidants. PhytoPs and PhytoFs represent relevant oxidative stress biomarkers in melon leaves. The use of natural antioxidants could reduce plant oxidative stress.

[Study on exogenous hormones inducing parthenocarpy fruit growth and development and quality of Siraitia grosvenorii].

To explore the growth and development and analyze the quality of the parthenocarpy fruit induced by exogenous hormones of Siraitia grosvenorii. the horizontal and vertical diameter, volume of the fruit were respectively measured by morphological and the content of endogenous hormones were determined by ELISA. The size and seed and content of mogrosides of mature fruit were determined. The results showed that the fruit of parthenocarpy was seedless and its growth and development is similar to the diploid fruit by hand pollination and triploid fruit by hand pollination or hormones. But the absolute value of horizontal and vertical diameter, volume of parthenocarpy fruit was less than those of fruit by hand pollination, while triploid was opposite. The content of IAA, ABA and ratio of ABA/GA was obviously wavy. At 0-30 d the content of IAA and ABA of parthenocarpy fruit first reduced then increased, content of IAA and GA parthenocarpy fruit was higher than that of fruit by hand pollination. Mogrosides of parthenocarpy fruit was close to pollination fruit. Hormones can induce S. grosvenorii parthenocarpy to get seedless fruit and the fruit shape and size and quality is close to normal diploid fruit by hand pollination and better than triploid fruit by hormone or hand pollination.

Enrichment of sugar content in melon fruits by hydrogen peroxide treatment.

Since sweetness is one of the most important qualities of many fruits, and since sugars are translocated from leaves to fruits, the present study investigates photosynthetic activity, activity of sugar metabolizing enzymes, sugar content in leaves and fruits and endogenous levels of hydrogen peroxide in leaves of melon plants treated with various dilutions of hydrogen peroxide, a nonspecific signaling molecule in abiotic stress. For this purpose, 4-month-old melon plants were treated with various concentrations (<50mM) of hydrogen peroxide by applying 300 mL per day to the soil of potted plants. The treatments resulted in increased fructose, glucose, sucrose and starch in the leaves and fruits. The most effective concentration of hydrogen peroxide was 20mM. During the day, soluble sugars in leaves were highest at 12:00 h and starch at 15:00 h. Furthermore, the peroxide treatment increased the photosynthetic activity and the activities of chloroplastic and cytosolic fructose-1,6-bisphosphatase, sucrose phosphate synthase and invertases. Thus, our data show that exogenous hydrogen peroxide, applied to the soil, can increase the soluble sugar content of melon fruits.

Citrus compost and its water extract for cultivation of melon plants in greenhouse nurseries. Evaluation of nutriactive and biocontrol effects.

Two different types of citrus composts, and their water extracts, were tested with regard to their utilisations as partial substitutes for peat in growing media for melon seedlings in greenhouse nurseries. Both compost showed higher plant growth than peat. Compost composed by citrus waste and green residue (C2) showed greater plant growth than compost obtained from the same organic matrices mentioned above further the addition of sludge obtained from citrus industry (C1). Compost C2 showed a greater auxinic effect than C1 and it was the only one that showed cytokinic effect. Both composts also demonstrated a biocontrol effect against Fusarium oxysporum for melon plants: the effects were also higher in C2 than in C1. Higher number of isolated fungi was active against F. oxysporum in compost C2, than compost C1. No different bacterial biocontrol efficacy was observed between both composts. The water extracts of both composts gave lower plant yields than their solid matrices, their relative effects being similar to those of the solid composts (C2 extract gave higher plant yields than the extract from C1). The biocontrol effects of compost water extracts followed the same trend.

The use of powder and essential oil of Cymbopogon citratus against mould deterioration and aflatoxin contamination of "egusi" melon seeds.

Experiments were carried out to determine the potential of using the powder and essential oil from dried ground leaves of Cymbopogon citratus (lemon grass) to control storage deterioration and aflatoxin contamination of melon seeds. Four mould species: Aspergillus flavus, A. niger, A. tamarii and Penicillium citrinum were inoculated in the form of conidia suspension (approx. 10(6) conidia per ml) unto shelled melon seeds. The powdered dry leaves and essential oil from lemon grass were mixed with the inoculated seeds at levels ranging from 1-10 g/100 g seeds and 0.1 to 1.0 ml/100 g seeds respectively. The ground leaves significantly reduced the extent of deterioration in melon seeds inoculated with different fungi compared to the untreated inoculated seeds. The essential oil at 0.1 and 0.25 ml/100 g seeds and ground leaves at 10 g/100 g seeds significantly reduced deterioration and aflatoxin production in shelled melon seeds inoculated with toxigenic A. flavus. At higher dosages (0.5 and 1.0 ml/100 g seeds), the essential oil completely prevented aflatoxin production. After 6 months in farmers' stores, unshelled melon seeds treated with 0.5 ml/ 100 g seeds of essential oil and 10 g/100 g seeds of powdered leaves of C. citratus had significantly lower proportion of visibly diseased seeds and Aspergillus spp. infestation levels and significantly higher seed germination compared to the untreated seeds. The oil content, free fatty acid and peroxide values in seeds protected with essential oil after 6 months did not significantly differ from the values in seeds before storage. The efficacy of the essential oil in preserving the quality of melon seeds in stores was statistically at par with that of fungicide (iprodione) treatment.

Agrobacterium-mediated transformation of bottle gourd (Lagenaria siceraria Standl.).

We describe a procedure for producing transgenic bottle gourd plants by inoculating cotyledon explants with Agrobacterium tumefaciens strain AGL1 that carries the binary vector pCAMBIA3301 containing a glufosinate ammonium-resistance (bar) gene and the beta-D-glucuronidase (GUS) reporter gene. The most effective bacterial infection was observed when cotyledon explants of 4-day-old seedlings were co-cultivated with Agrobacterium for 6-8 days on co-cultivation medium supplemented with 0.1-0.001 mg/l L-alpha-(2-aminoethoxyvinyl) glycine (AVG). The putatively transformed shoots directly emerged at the proximal end of cotyledon explants after 2-3 weeks of culturing on selection medium containing 2 mg/l DL-phosphinothricin. These shoots were rooted after 3 weeks of culturing on half-strength MS medium containing 0.1 mg/l indole acetic acid and 1 mg/l DL-phosphinothricin. Transgenic plants were obtained at frequencies of 1.9%. Stable integration and transmission of the transgenes in T1 generation plants were confirmed by a histochemical GUS assay, polymerase chain reaction and Southern blot analyses. Genetic segregation analysis of T1 progenies showed that transgenes were inherited in a Mendelian fashion. To our knowledge, this study is the first to show Agrobacterium-mediated transformation in bottle gourd.

Expression of CycD3 is transiently increased by pollination and N-(2-chloro-4-pyridyl)-N'-phenylurea in ovaries of Lagenaria leucantha.

Lagenaria leucantha is an important vegetable crop and a potential model for the study of fruit development. To study the function of D cyclins in fruit development, full-length cDNA clones for two D cyclin genes were isolated from young ovaries of Lagenaria leucantha. They were classified as D3 cyclins by sequence similarities and phylogenetic analysis, and nominated LlCycD3;1 and LlCycD3;2, respectively. The deduced amino acid sequence of both LlCycD3 genes contained a retinoblastoma-binding motif and a PEST-destruction motif. Unpollinated ovaries failed to develop and eventually aborted. N-(2-chloro-4-pyridyl)-N'-phenylurea (CPPU) induced parthenocarpic fruit significantly larger than pollinated ones. In unpollinated ovaries, the expression of both LlCycD3 genes was abundant at anthesis and then suddenly decreased, concomitant with the cessation of cell division. Pollination/fertilization induced an activation of the cell cycle accompanied by a large increase in the transcript levels of LlCycD3;1 and LlCycD3;2 in young fruits. Treating ovaries with CPPU also reactivated cell division and transcription of CycD3 genes and the effect was more rapid and pronounced than after pollination/fertilization.