Functional characterization and expression analysis of cucumber (Cucumis sativus L.) hexose transporters, involving carbohydrate partitioning and phloem unloading in sink tissues.

Many hexose transporters (HTs) have been reported to play roles in sucrose-transporting plants. However, little information about roles of HTs in RFOs (raffinose family oligosaccharides)-transporting plants has been reported. Here, three hexose transporters (CsHT2, CsHT3, and CsHT4) were cloned from Cucumis sativus L. Heterologous expression in yeast demonstrated that CsHT3 transported glucose, galactose and mannose, with a K(m) of 131.9 µM for glucose, and CsHT4 only transported galactose, while CsHT2 was non-functional. Both CsHT3 and CsHT4 were targeted to the plasma membrane of cucumber protoplasts. Spatio-temporal expression indicated that transcript level of CsHT3 was much higher than that of CsHT2 and CsHT4 in most tissues, especially in peduncles and fruit tissues containing vascular bundles. GUS staining of CsHT3-promoter-ß-glucuronidase (GUS) transgenic Arabidopsis plants revealed CsHT3 expression in tissues with high metabolic turnover, suggesting that CsHT3 is involved in sugar competition among different sink organs during plant development. The transcript levels of CsHT3 and cell wall invertase genes increased in peduncles and fruit tissues along with cucumber fruit enlargement, and CsHT3 localized to phloem tissues by immunohistochemical localization; These results suggest that CsHT3 probably plays an important role in apoplastic phloem unloading of cucumber fruit.

Molecular cloning, characteristics and low temperature response of raffinose synthase gene in Cucumis sativus L.

Raffinose synthase (RS, EC2.4.1.82) is one of the key enzymes that channels sucrose into the raffinose family oligosaccharides (RFOs) biosynthetic pathway. However, the gene encoding RS is poorly characterized in cucumber (Cucumis sativus L.), which is a typical RFOs-translocating plant species. Here we isolated the gene encoding RS (CsRS) from the leaves of cucumber plants. The complete cDNA of CsRS consisted of 2552 nucleotides with an open reading frame encoding a polypeptide of 784 amino acid residues. Reverse transcription-polymerase chain reaction and RNA hybridization analysis revealed that expression of CsRS was the highest in leaves followed by roots, fruits, and stems. The RS activity was up-regulated and the raffinose content was high in the leaves of transgenic tobacco with over-expression of CsRS, while both the RS activity and the raffinose content decreased in the transgenic cucumber plants with anti-sense expression of CsRS. The expression of CsRS could be induced by low temperature and exogenous phytohormone abscisic acid (ABA). In cucumber growing under low temperature stress, CsRS expression, RS activity and raffinose content increased gradually in the leaves, the fruits, the stems and the roots. The most notable increase was observed in the leaves. Similarly, the expression of CsRS was induced in cucumber leaves and fruits with 200 µM and 150 µM ABA treatments, respectively.

[Effects of alternative furrow irrigation and nitrogen application rate on photosynthesis, growth, and yield of cucumber in solar greenhouse].

This paper studied the effects of alternative furrow irrigation and nitrogen (N) application rate (no N, optimal N, and conventional N) on the photosynthesis, growth characteristics, yield formation, and fruit quality of cucumber (Cucumis sativus) cultivar Jinyu No. 5 in a solar greenhouse in winter-spring growth season and autumn-winter season. Under alternative furrow irrigation, the net photosynthetic rate of upper, middle, eand lower leaves was appreciably lower and the transpiration rate decreased significantly, and the transient water use efficiency of upper and middle leaves improved, as compared with those under conventional irrigation. Stomatal factor was the limiting factor of photosynthesis under alternative furrow irrigation. The photosynthesis and transient water use efficiency of functional leaves under alternative furrow irrigation increased with increasing N application rate. Comparing with conventional irrigation, alternative furrow irrigation decreased leaf chlorophyll content and plant biomass, but increased root biomass, root/shoot ratio, and dry matter allocation in root and fruit. The economic output under alternative furrow irrigation was nearly the same as that under conventional irrigation, whereas the water use efficiency for economic yield increased significantly, suggesting the beneficial effects of alternative furrow irrigation on root development and fruit formation. With the increase of N application rate, the leaf chlorophyll content, chlorophyll a/b, specific leaf mass, plant biomass, economic yield, and fruit Vc and soluble sugar contents under alternative furrow irrigation increased, but no significant difference was observed between the treatments optimal N and conventional N. N application had little effects on the water use efficiency for economic yield. The economic yield and biomass production of the cucumber were significantly higher in winter-spring growth season than in autumn-winter growth season.

[Effects of temperature regime on low-light tolerance of Cucumis sativus seedling leaves in their photosynthesis].

In a phytotron, the effects of three temperature regimes (day/night 25 degrees C/18 degrees C, optimal temperature; 15 degrees C/9 degrees C, suboptimal temperature; and 9 degrees C/7 degrees C, low temperature) on the low-light (75-85 micromol x m(-2) x s(-1)) tolerance of two Cucumis sativus cultivars (shade-susceptible Jinyan 2 and shade-tolerant Deltastar) seedling leaves in their photosynthesis were studied. The results showed that under low light, the SPAD, net photosynthesis rate (Pn), stomatal conductance (Gs), transpiration rate (Tr), water use efficiency (WUE), actual photochemical efficiency of PS II in the light (phi(PS II)), and photochemical quenching of chlorophyll fluorescence (q(p)) of cucumber leaves decreased, with the decrement getting more with decreasing temperature, while the superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) activities were in adverse. During the recovery process after low-light stress relieved, the parameters of gas exchange and chlorophyll fluorescence of the leaves recovered gradually, and the recovery of some gas exchange parameters lagged to that of chlorophyll fluorescence parameters. Under low light, the lower the temperature, the more damage the photosynthesis apparatus suffered, and the lesser tolerance to low light the cucumber leaves had in their photosynthesis. During the low temperature and low light treatment period, the decrease of Pn, phi(PS II), and q(p) was more obvious for Jinyan 2 than for Deltastar; and during the relief period, the recovery of these parameters was slower for Jinyan 2 than for Deltastar. It was indicated that Jinyan 2 had weaker tolerance to low temperature and/or low light in its photosynthesis than Deltastar.